--- Log opened Mon Jul 13 00:00:09 2015 00:57 -!- justanotheruser [~Justan@unaffiliated/justanotheruser] has quit [Ping timeout: 250 seconds] 01:56 -!- Porb_ [~Porbus@CPE-124-176-219-129.lns5.win.bigpond.net.au] has quit [Read error: Connection reset by peer] 02:33 -!- Viper168_ [~Viper@unaffiliated/viper168] has joined ##hplusroadmap 02:35 -!- Viper168 [~Viper@unaffiliated/viper168] has quit [Ping timeout: 250 seconds] 02:37 -!- Viper168_ is now known as Viper168 02:55 -!- chris_99 [~chris_99@unaffiliated/chris-99/x-3062929] has joined ##hplusroadmap 03:08 -!- justanotheruser [~Justan@unaffiliated/justanotheruser] has joined ##hplusroadmap 03:24 -!- soylentbomb [~k@unaffiliated/soylentbomb] has quit [Quit: leaving] 03:32 -!- zadock [~outsider@cthulhu.tuiasi.ro] has joined ##hplusroadmap 03:34 -!- zadock [~outsider@cthulhu.tuiasi.ro] has quit [Remote host closed the connection] 03:38 -!- PatrickRobotham [uid18270@gateway/web/irccloud.com/x-vqbxlinqdnrfvdvb] has quit [Quit: Connection closed for inactivity] 03:43 -!- drewbot_ [~cinch@ec2-54-226-221-122.compute-1.amazonaws.com] has joined ##hplusroadmap 03:49 -!- drewbot [~cinch@ec2-54-211-61-160.compute-1.amazonaws.com] has quit [Ping timeout: 256 seconds] 03:58 -!- knobuddy is now known as ghostcaptain 04:06 -!- PatrickRobotham [uid18270@gateway/web/irccloud.com/x-duyxyyetgupmyrla] has joined ##hplusroadmap 04:22 -!- chris_99 [~chris_99@unaffiliated/chris-99/x-3062929] has quit [Quit: Ex-Chat] 04:23 -!- chris_99 [~chris_99@unaffiliated/chris-99/x-3062929] has joined ##hplusroadmap 05:30 < dpk> .privacy 05:30 < yoleaux> dpk: This channel is public. When I am asked when I last saw you, I may repeat things you say and what time it was when you said them. 05:30 < ghostcaptain> ʘ_ʘ 05:33 -!- Porb [~Porbus@CPE-124-176-219-129.lns5.win.bigpond.net.au] has joined ##hplusroadmap 05:34 < fenn> ghostcaptain: if that was a surprise, may i humbly submit that you please read the channel topic 05:40 -!- Porb [~Porbus@CPE-124-176-219-129.lns5.win.bigpond.net.au] has quit [Ping timeout: 265 seconds] 05:41 -!- Porb [~Porbus@CPE-124-176-219-129.lns5.win.bigpond.net.au] has joined ##hplusroadmap 05:46 -!- eudoxia [~eudoxia@r167-56-36-161.dialup.adsl.anteldata.net.uy] has joined ##hplusroadmap 06:01 < ghostcaptain> that topic is a mess 06:01 < ghostcaptain> plus i forgot 06:01 < ghostcaptain> ^_^ 06:02 < streety> probably best to assume every channel you are on is logged by someone 06:03 < fenn> best to assume all traffic everywhere is logged by someone 06:03 < chris_99> mmm 06:03 < fenn> but this channel is google-searchable 06:04 < ghostcaptain> i know i'm logging 06:05 < streety> poorly searchable 06:06 < fenn> if you want better searchability you can do this and then grep: 06:06 < fenn> wget -r -np -nc --exclude-dir="logs/html/,logs/hplusroadmap-logs/" --reject "2008-03-25.log" gnusha.org/logs/ 06:06 < chris_99> heh, why reject that one out of interest? 06:06 < fenn> because it changes every time and it's dumb to download it when it's just a duplicate of the individual log files 06:07 < chris_99> ah 06:10 < streety> thanks 06:10 < fenn> note that this will give a bad file for the day you downloaded it because -nc doesn't try to re-download files that exist (use -c for that, which is slower) 06:11 < fenn> i mean the next time you download logs, the "head" from last time will be incomplete 06:11 < fenn> so you could just delete the last log file before downloading 06:15 -!- ebowden [~ebowden@101.177.244.210] has quit [Remote host closed the connection] 06:17 -!- proofoflogic [uid65184@gateway/web/irccloud.com/x-trgyjteenmtzpizq] has quit [Quit: Connection closed for inactivity] 06:33 -!- nottimschmidt [~timschmid@35.10.220.83] has joined ##hplusroadmap 06:34 -!- nottimschmidt [~timschmid@35.10.220.83] has left ##hplusroadmap [] 06:35 -!- nottimschmidt [~timschmid@35.10.220.83] has joined ##hplusroadmap 06:35 -!- nottimschmidt [~timschmid@35.10.220.83] has quit [Client Quit] 06:35 -!- nottimschmidt [~timschmid@35.10.220.83] has joined ##hplusroadmap 06:36 -!- Madplatypus [uid19957@gateway/web/irccloud.com/x-ckarasonpysmdemc] has quit [Quit: Connection closed for inactivity] 06:46 -!- ebowden [~ebowden@101.177.244.210] has joined ##hplusroadmap 06:48 < kanzure> i guess we could throw up a search form and an index, like solr or something 06:48 < kanzure> but then i would feel bad about not indexing special characters 06:51 < kanzure> CaptHindsight: airlocks are important 06:52 -!- ebowden [~ebowden@101.177.244.210] has quit [Ping timeout: 248 seconds] 06:53 < kanzure> so who here wants to house the dna synthesizer once it's complete? any takers. 06:55 -!- JayDugger [~jwdugger@108.19.186.58] has joined ##hplusroadmap 07:04 < ParahSailin> this one labs quote? 07:05 < kanzure> yes 07:06 < kanzure> also- i think the air jet dryer thingy will blow away the dna unless it's tied to the surface. so that's more chemistry that needs to be figured out. 07:06 < ParahSailin> what likelihood would you assign to it working 07:06 < kanzure> motion control elements, 100% likely 07:06 < kanzure> 100 bp strands per spot, no clue.. 07:06 < fenn> mumble mumble acetonitrile toxicity flammability 07:07 < fenn> mumble large heavy fragile expensive object that needs to be treated carefully 07:07 < kanzure> well the primary reason for the size is the number of spots, i think 07:08 < kanzure> oh plus whatever form factor we can purchase argon in 07:08 < ParahSailin> 60cm travel in one dimension? 07:08 < fenn> the size is mostly because it's made on an optical breadboard i think 07:09 < kanzure> this isn't going to happen if i can't find somewhere to put it 07:10 < ParahSailin> can put it here 07:10 < fenn> going once 07:10 < fenn> twice 07:10 * nottimschmidt is asking about storing it in lab space at Michigan State University and Washington University. 07:11 < nottimschmidt> Genspace might be interested in NYC 07:11 < kanzure> genspace hates us 07:11 < nottimschmidt> I see :-/ 07:11 < kanzure> but nice idea 07:12 < kanzure> posam (another inkjet dna microarrayer thingy) is actually housed in washington in the hood laboratory 07:13 < kanzure> https://www.systemsbiology.org/bio/leroy-hood/ 07:13 < kanzure> http://techdev.systemsbiology.net/posam/ 07:13 < nottimschmidt> I know a postdoc there, in the Kerr and Klavins labs 07:13 < ParahSailin> you going with this instead of cmos because its cheaper? 07:13 < nottimschmidt> https://twitter.com/LuisZaman 07:13 < kanzure> when you say cmos what do you mean 07:13 < nottimschmidt> He does 3D printing and microfluidics stuff 07:14 < ParahSailin> electrochemical array 07:14 < kanzure> i believe the electrochemical cmos array thingy still used inkjet stuff- it just had electrodes at each spot for generating acid or something 07:14 < ParahSailin> the thing azco sells? 07:15 < ParahSailin> pretty sure no inkjets involved 07:15 < kanzure> oh was this the light valve one? with LCDs or DMDs? 07:16 < fenn> there was a paper about photoelectrochemical synthesis and another one about just electrochemical synthesis 07:16 < fenn> they used the same reaction mechanism 07:17 < ParahSailin> CustomArray 07:17 < ParahSailin> no optics, just electronics 07:18 < kanzure> "My only other comments are that this looks like a great V1 machine. LinuxCNC as a requirement is a barrier to entry - because it necessitates a dedicated control computer. And a reduced-cost and reduced-wiring-complexity v2 might strive toward single-board electronics" 07:18 < kanzure> "FPGAs are similarly a barrier to entry, due to the required non-free tooling, and increased cost. But those barriers are actively being removed at the moment, at least for certain FPGAs. 07:18 < kanzure> https://wiki.debian.org/FPGA " 07:18 < kanzure> ParahSailin: i wasn't aware that they were using *only* electrodes to control the chemistry. how do they isolate which strands get capped/uncapped? 07:19 < fenn> the benefit of FPGA is higher performance/throughput and it enables servo motors instead of stepper motors 07:19 < ParahSailin> addressing each electrode individually 07:19 < kanzure> but.. so .. the claim is that each electrode is able to control the capping/uncapping of a long oligo? 07:20 < ParahSailin> its not a claim, its been a commercial product for almost 10 years 07:20 < fenn> i'm not sure either of these features are necessary for a proof of concept, but it's not a super huge cost relative to the overall quoted cost of the system 07:20 < ParahSailin> you can even order a test run from azco for a couple k, to see if the error rate is good enough 07:20 < nottimschmidt> fenn: servos can be done in software, on CPUs too. LinuxCNC does that, for instance. Nothing special about FPGAs enables them. 07:21 < fenn> nottimschmidt: no, in fact you will not get good results doing servo control in software without some kind of dedicated interface card to watch the optical encoders 07:21 < nottimschmidt> In CPUs, these are called "external interrupts" 07:22 < fenn> even a dedicated microcontroller can not keep up with a servo motor with common encoder resolutions and speeds 07:22 < fenn> there is usually some kind of hardware counter to add up the encoder transitions since last interrupt 07:23 < nottimschmidt> One servo per, little interrupt handler to dispatch control I/O, bob's your uncle. You want to be running a realtime OS, but something like http://bertos.org/ manages this on AVR, ARM, and x86 with a sane interface. 07:23 < ParahSailin> even less than a couple k https://azcobiotech.com/oligo-array-12k-pool-10-to-130-mer-in-length-80-0012.html 07:23 < fenn> that only works for low resolutions and/or slow speeds, which negates the point in using servo motors in the first place 07:23 < nottimschmidt> Certainly speed is affected by the choice of solution. Have you done the math on what speed and resolution you need? 07:24 < fenn> modern PC CPUs are terrible at realtime interrupts 07:24 < kanzure> ParahSailin: eventually i want to look into the physics or chemistry of that reaction mechanism, because i was not aware that it was electrode-only control.... 07:24 < ParahSailin> isnt that just an os issue? 07:24 < fenn> mumble mumble pipeline context switch mumble 07:25 < ParahSailin> i think its this http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1762329/ 07:25 < kanzure> nottimschmidt: well, we would like to have millions of spots- which would get us pretty close to the size of a genome 07:25 < fenn> i don't know the specifics at a low level but i do know that people more experienced than me all agree that it's impossible 07:25 < kanzure> .title 07:25 -!- Porb [~Porbus@CPE-124-176-219-129.lns5.win.bigpond.net.au] has quit [Quit: Leaving] 07:25 < yoleaux> Electrochemically Generated Acid and Its Containment to 100 Micron Reaction Areas for the Production of DNA Microarrays 07:25 -!- nsh [~lol@wikipedia/nsh] has quit [Ping timeout: 246 seconds] 07:25 < ParahSailin> ask a cmos custom prototyper for a quote on that apparatus 07:26 < nottimschmidt> fenn: I've written / heavily modified some (admittedly slow, at 16Mhz) realtime motor control interrupt handlers. Seems very possible to me. 07:26 < kanzure> i guess fenn is right, here, i was probably mistakening this for the "photolithography dna synthesis" method that used photogenerated acid or something for one of the steps, or electrode-generated acid for one of the steps. 07:26 < kanzure> ParahSailin: how are the strands tied to the electrode? 07:26 < fenn> nottimschmidt: was 16MHz the speed of the cpu or the speed of the encoder transitions? 07:27 < ParahSailin> photolithography with chrome masks is how affy is stuck doing it 07:27 < nottimschmidt> fenn: cpu speed 07:27 < kanzure> ParahSailin: *masked* photolithography! ouch. what the fuck? 07:27 < fenn> nottimschmidt: roughly how fast did the encoders go? 07:28 < nottimschmidt> they topped out around 40,000 pulses per second 07:28 < kanzure> not bad; i think this piezo printhead does 16 kHz drops 07:29 < ParahSailin> cleavable linker http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1762329/figure/pone-0000034-g006/ 07:29 -!- nsh [~lol@wikipedia/nsh] has joined ##hplusroadmap 07:29 < nottimschmidt> I think you could do a lot more with a 100Mhz ARM M3 let alone something cell-phone class. 07:29 < kanzure> right... but i mean the initial placement of the linker. 07:29 < fenn> nottimschmidt: ok that's about the max rate for outputting stepper motor pulses from a parallel port 07:30 < nottimschmidt> yeah, parallel ports are not great 07:30 -!- nottimschmidt [~timschmid@35.10.220.83] has quit [Remote host closed the connection] 07:30 < kanzure> he.. died? 07:30 < fenn> not sure what just happened 07:32 < kanzure> so wouldn't it be better to isolate each reaction in a drop instead ofj ust hoping the acids don't brownian motion themselves over to the other electrode areas 07:32 < ParahSailin> some sort of a reactive chemical reagent? 07:33 < ParahSailin> kanzure: which is why i suggest spending the $1650 on an azco array run, and then a few k more on a nextseq run to see how the error rate is 07:34 < fenn> can't you just ask them what the error rate is 07:34 < fenn> surely someone has quantified this already 07:34 < ParahSailin> well, the application for these is generally just as hybridization probes 07:35 < kanzure> i also wonder if individual dna could be harvested from that approach (oops i mean specific sequences); because i think it's all in the same water bath 07:35 < ParahSailin> like stick biotin on your whole pool and do X 07:35 < kanzure> or er, not water... 07:35 < fenn> since they're bound to the surface (right?) you could 3d print wells around them or something, assuming they give you the plate 07:36 < ParahSailin> "In contrast, in the electrochemical case reagent generated by the electrode is able to diffuse into the PRL but is prevented from excessive diffusion by the presence of a buffering agent in the surrounding solution." 07:36 < kanzure> then how does it migrate to the tip of the oligo? 07:36 < ParahSailin> so i think they figured out a solution 07:36 < ParahSailin> " Near the electrode, the buffer is overwhelmed, allowing reaction of the ECG reagent with the PRL bound substrate; but further away, the concentration of the diffusing reagent is insufficient to overcome the concentration of the buffering agent, and no reaction occurs." 07:37 < fenn> the oligo is only ... (0.2nm * number of base pairs) long 07:37 < kanzure> i suppose the oligo is very short- oh fenn is on it 07:37 < fenn> i think stereolithography/DLP 3d printing works the same way 07:37 -!- nottimschmidt [~timschmid@35.10.220.177] has joined ##hplusroadmap 07:37 < fenn> wb nottimschmidt 07:38 < nottimschmidt> thanks, sorry, power adapter issues 07:38 < ParahSailin> get a bouncer 07:38 < nottimschmidt> Will do 07:39 < ParahSailin> microsoft gives away $750/mo in azure to practically anyone 07:39 < nottimschmidt> I've got a cloudatcost machine already running some things. Will set it up there. 07:41 < kanzure> so i agree that an electrode microarray would be a good and sane thing to work with in the near future 07:41 < kanzure> although, an inkjet plus motion control is also useful for other reasons 07:41 < kanzure> and can possibly be coerced into higher densities 07:42 < nottimschmidt> So anyway, FPGAs can be a great solution. http://papilio.cc/ and other low-cost FPGA boards have been starting to grow a larger community of knowledgeable folk. And they can run AVR and ARM soft cores for offloading some of the realtime processing, just like a microcontroller. 07:43 < nottimschmidt> I'm really happy to see some Free tools start to show up for some of these boards. 07:43 < nottimschmidt> They're still early days, though 07:43 < ParahSailin> kanzure: it seems like you have a bias to have something that anyone can build themselves at home, whereas you should go with what is actually the viable technology 07:44 < CaptHindsight> the whole PC vs uController thing will cost you more money, there isn't a single board microcontroller board that can handle the closed loop and printhead control on the market 07:44 < CaptHindsight> you'll have to reinvent that wheel 07:44 < kanzure> CaptHindsight: i misunderstood the electrode acid approach, it's a microelectrode array and no LCD or DMD or inkjet 07:45 < nottimschmidt> CaptHindsight: if I can do it (with some speed limitations) on a 16Mhz AVR (and I have), what makes you say that no uC board on the market can do it? 07:46 < kanzure> ParahSailin: well i'm biased towards the inkjet design because we have someone that is willing to build it 07:46 < CaptHindsight> nottimschmidt: I know that the Linuxcnc works well. Try and do it with your AVR 07:46 < ParahSailin> kanzure: and thats only because you emailed them before you emailed a cmos prototype shop 07:47 < nottimschmidt> CaptHindsight: I hack CNC firmwares for the AVR. 07:48 < kanzure> nottimschmidt: what's wrong with linuxcnc anyway? your argument is that it's "bulky" right? 07:48 < CaptHindsight> kanzure: have a link to clarify the "electrode acid approach"? 07:49 < ParahSailin> scroll 07:50 < CaptHindsight> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1762329/ ? lots of noise in here today 07:51 < nottimschmidt> kanzure: it shoves a lot of the realtime work onto the big desktop CPU, precisely where we all agree it shouldn't be. That necessitates running a realtime OS on a machine you interact directly with, which means external interrupts are being triggered not just by the machine, but by the unpredictable user. The whole thing requires additional hardware over a uC or self-hosted FPGA-based solution, setup is more complex, ladder logic isn't wi 07:51 < nottimschmidt> dely understood, etc etc. 07:52 < nottimschmidt> LinuxCNC works and is very flexible. Hence my comment about this design being a good v1 machine. 07:54 < juri_> ok, i've caught up. 07:55 < juri_> I'd be coordinating a batch of propeller MCUs, but the FPGA chosen for this project is my personal favorite. 07:56 < juri_> it's supported by a free software toolchain, as well. we could load a MCU into it, and make it appear to the PC as a serial port accepting GCODE. 07:57 < nottimschmidt> juri_: that's awesome 07:57 < juri_> we could even use pronterface for delivering the gcode. 07:58 < CaptHindsight> I thought we'd go the electrode acid approach next 07:58 < nottimschmidt> So long as Free tools work well with it, and cost isn't multiple orders of magnitude different, the FPGA sounds like the best solution. 07:58 < juri_> linuxcnc might get us across the finish line faster, if we approach the problem from the right angle, break it into jobs we can test seperately, and apply all the brainpower we have. 07:58 < nottimschmidt> Heat and power consumption aren't likely to be issues 08:00 < CaptHindsight> Linuxcnc is guaranteed to work, then you can't use the working example to make whatever else you'd like 08:01 < CaptHindsight> BBB, smoothie, parallel propellers, stm32 etc etc 08:01 < CaptHindsight> sorry can't/can 08:01 < nottimschmidt> juri_: I'm not wedded to gcode either, there are other machine control languages like OpenSBP, and I'm sure some genomics-specific domain specific languages. Open to ideas. 08:02 < juri_> I think gcode is the right tool. it's just a question of how you use it. 08:02 < nottimschmidt> k 08:02 < juri_> Honestly, i think you should be modeling the machine more like a 3d printer in how you speak to it. 08:03 < juri_> this makes the "slicer" more complicated, but removes a lot of the realtime interaction's complexity during the debugging stage. 08:04 < ParahSailin> kanzure: and if you figured out how to electrochemically cleave the linker, then you have a way to suck up individual oligos 08:04 < nottimschmidt> Yes. Doing as much work ahead of time as possible, in the "slicer" is an excellent idea. 08:04 < CaptHindsight> I think we should make more tools so that people can easily make things like the CMOS array 08:05 < juri_> I don't think the gcode interpreter should have any idea what the (x,y) offsets of the nozzles are. 08:06 < juri_> we should treat them as extruders. "go to position XYZ" "go from that position to this new position while firing a dropplet from nozzles (42-85)"... etc. 08:07 < juri_> complicated "slicer", but simple gcode interpreter. 08:07 < kanzure> i'm confused about the eat-gcode-by-serial-port thing, is it the fpga that is interpreting the gcode? why... gcode interpreters suck, why not just interpret it before sending data to the equipment? 08:07 < fenn> in the current design the gcode doesn't say anything about nozzles at all, that's handled by a preprocessor (that has yet to be written) 08:07 < kanzure> yes i'm not sure the optimal gcode modification method here. still looking at linuxcnc docs. 08:07 < kanzure> i understand the HAL stuff, but not how it interfaces with gcode stuff. 08:08 < kanzure> ParahSailin: we can still use electrode arrays with this approach 08:08 < juri_> kanzure: that's prospective, and down the line. i think implementing in linuxcnc is probably faster. 08:08 < nottimschmidt> kanzure: the FPGA would do the interpreting, yes. Reason is that the machine control signals happen in real-time which desktops suck at. 08:08 < nottimschmidt> Desktops want to do large batch job type things. 08:08 < ParahSailin> sounds like a lot of reduplicated effort 08:08 < kanzure> nottimschmidt: so who is going to write the vhdl for a gcode interpreter? isn't that a fate worse than death 08:08 < fenn> i'm thinking of it like the print head nozzle firing is slaved to the movement; the fpga fires the nozzles at the correct position but otherwise has no control over the machine 08:09 < nottimschmidt> kanzure: soft-AVR core on the FPGA w/ a firmware? 08:09 < CaptHindsight> fenn: thats the approach I'd recommend 08:09 < juri_> kanzure: you don't. you write a gcode interpreter for an MCU, then upload that MCU to the fpga.. again, in an ideal world. ;) 08:09 < kanzure> so the fpga is not moving the gantry. ok. 08:09 < CaptHindsight> that is how most inkjet printers operate 08:09 < nottimschmidt> kanzure: it's better than it sounds. Opencores has a 100Mhz soft-avr core. 08:10 < kanzure> nottimschmidt: i have not looked at opencores in a while, that's neat. 08:10 -!- ThomasEgi [~thomas@panda3d/ThomasEgi] has quit [Ping timeout: 252 seconds] 08:10 < juri_> nottimschmidt: the propellers get 200MIPS at 32Mhz. 8 core microcontroller. crazy design. ;) 08:10 < nottimschmidt> Yes. I love the prop. 08:10 < kanzure> ParahSailin: you still need nozzles, you still need a neutral atmosphere, it's a lot of similar equipment 08:11 < fenn> it's sort of hard to talk about because the fpga is performing multiple functions in parallel; it's also watching the servo encoders and generating motor control signals etc 08:11 < nottimschmidt> juri_: also love that it was designed for production on metalized gate array. 08:11 < kanzure> fenn: ah i see. 08:11 < ParahSailin> kanzure: actually you dont even need to go to a cmos place https://azcobiotech.com/oligo-array-12k-chip-cleavable-19-5012.html 08:11 < ParahSailin> just buy it and breadboard some shit around it 08:11 < kanzure> isn't this the same link 08:12 < ParahSailin> nah, this is a "consumable" cmos array 08:12 < fenn> the thing that fires the nozzles can see what position the motors are at, but has no control over them 08:12 < ParahSailin> this is much cheaper than the quote you got 08:12 < fenn> but this "thing" only exists as a pattern of activated logic gate cells 08:12 < CaptHindsight> yes the printhead just fires as it get gated encoder pulses 08:13 < kanzure> ParahSailin: btw CaptHindsight made the quote 08:13 < juri_> CaptHindsight: you're not wanting to control motion from the same FPGA? 08:13 < CaptHindsight> yeah, we looked at those 12k chips last week 08:14 < fenn> FPGA only does motion interpolation at a low level 08:14 < fenn> like linear interpolation (maybe not even circular interpolation) 08:14 < kanzure> ParahSailin: you make some good arguments. 08:15 < CaptHindsight> juri_: I want it to work in a few weeks. I'm not spending any time on reinventing the wheel on the first rev 08:15 < juri_> CaptHindsight: so, describe the motion control to me. 08:16 < ParahSailin> kanzure: so the steps i would do is order a run done on one of these customarray chips, send it to ucdavis to run on a miseq for another 1500, then if satisfactory error rate, get a 12k chip and design electronics and fluid handling so you dont have to buy the official 300k machine 08:16 < CaptHindsight> juri_: if you want to spend the time stuffing it all the FPGA you will be able to 08:16 < CaptHindsight> it's an open design 08:16 < juri_> because i'm really concerned that if you try to expose the 'blast a bitmap' code to the CPU in realtime, you'll lose those weeks to testing and defects. 08:17 < kanzure> ParahSailin: send the array to ucdavis? 08:17 < ParahSailin> any cal students could get a $500 discount from davis it looks like 08:17 < kanzure> or send the cleaved oligos 08:17 < ParahSailin> kanzure: cleaved oligos 08:17 < kanzure> this seems to claim to have a cleaving function. hm. 08:17 < CaptHindsight> juri_: the FPGA on the Mesa card is about 10% full with the 3 servos 08:17 < ParahSailin> azco will send a tube of pooled, cleaved oligos 08:18 < kanzure> oh 08:18 < ParahSailin> you just pass it on to davis 08:18 < CaptHindsight> so we have enough room left to control the printhead with some buffering 08:18 < fenn> CaptHindsight: it's been a while and i forget the details.. does mesa's hotmot thing do PID on the FPGA or in the PC CPU? 08:18 < CaptHindsight> it's how we have been making printhead controllers for years 08:18 < fenn> hostmot* 08:18 < CaptHindsight> with the FPGA it's in the FPGA 08:18 < juri_> CaptHindsight: ah, that's fine. 08:19 -!- nmz787_i [ntmccork@nat/intel/x-nxxygqsseduszlkg] has joined ##hplusroadmap 08:19 < CaptHindsight> juri_: if we only had 10% of the gates left I wouldn't recommend this or what to spend the time hand packing gates 08:20 < CaptHindsight> what/want 08:20 < juri_> CaptHindsight: if you run out, you can always just add another FPGA board, and tie the I/O between the two boards. 08:21 < juri_> that makes it a compiler problem, nothing more. 08:21 < nmz787_i> CaptHindsight: I saw your quote, you listed solenoids... but I couldn't tell what for... what are they hooked up to? 08:21 < CaptHindsight> we have large FPGA cards that are Linuxcnc compatible and we also can plug in multiple FPGA cards 08:22 < juri_> CaptHindsight: i like this model. stick to it. ;) 08:22 < CaptHindsight> I have systems with 7 PCIe FPGA cards on one motherboard 08:22 -!- ThomasEgi [~thomas@panda3d/ThomasEgi] has joined ##hplusroadmap 08:22 < CaptHindsight> running multiple printheads with 100x the Epson nozzles 08:23 < juri_> Impressive. 08:23 < nottimschmidt> yeah, very cool 08:24 < CaptHindsight> http://global.kyocera.com/prdct/printing-devices/inkjet-printheads/ think about trying to supply data to 16 or 32 of these 08:24 < fenn> how do you get the bitmap data into the fpga at the right time? 08:24 < kanzure> HAL 08:24 < fenn> presumably there's some sort of buffer somewhere 08:24 < juri_> I seriously reccomend you just encode it into the gcode. 08:24 < CaptHindsight> >100 million drops per printhead per second 08:24 < juri_> it will make your world much easier to debuh. 08:24 < fenn> i recommend against that because of how linuxcnc passes g-code around internally 08:25 -!- ghostcaptain [~IceChat9@unaffiliated/knobuddy] has quit [Remote host closed the connection] 08:25 < juri_> fenn: howso? 08:25 < kanzure> the one major argument i have against not-in-the-gcode is that then you're not /really/ using gcode to talk with the machine. but this is only a weak argument. 08:26 < juri_> it'll make simulation harder. 08:26 < juri_> and debugging. 08:26 < kanzure> fenn: i think the fpga reports encoder stuff back to the pc 08:26 < fenn> well it generates a message for each g-code command and puts it on a queue and then something else takes things off the queue and looks at them and determines their C++ type in order to figure out what kind of message it is and then passes it to another thing that figures out what to do with this particular message type... it ends up generating a lot of work if you do it literally a million times 08:26 < fenn> a second 08:26 < kanzure> yes i noticed that linuxcnc seems to pass around gcode by stdout? it was amusing 08:26 < nottimschmidt> I implemented fast (in the interrupt handler) bitmap raster support for lasers in Marlin, and encoded the bitmap data into the gcode. Works a treat. Single "nozzle" though. 08:27 < fenn> kanzure: if it's using stdout it's not technically part of emc but rather some preprocessor script 08:27 < kanzure> ah okay 08:27 < CaptHindsight> http://wiki.linuxcnc.org/cgi-bin/wiki.pl?EMC_Components Linuxcnc with block diagrams of how it works 08:27 < ParahSailin> i give up 08:27 < nottimschmidt> we base64-endoded the bitmap data, and shoveled it in with a custom G code. 08:28 < juri_> not a bad approach. 08:28 < fenn> ParahSailin: what are you giving up? 08:29 < nmz787_i> nottimschmidt: x86 I/O that isn't pipelined is extremely laggy 08:29 < nottimschmidt> nmz787_i: yep 08:29 < CaptHindsight> using the FPGA's we can do the PID in the FPGA, there are Ethernet FPGA cards that only need a 1mS loop vs few uS loop in the PC 08:30 < CaptHindsight> and do it over ethernet 08:30 < kanzure> fenn: he's giving up on trying to convince me to use a microelectrode array 08:30 < nottimschmidt> nmz787_i: deep caches, pipelines, competing threads, boatloads of I/O, and non-realtime OSes all contribute to this problem. That's why I prefer to offload g-code processing to a uC or soft-core in an FPGA running in real time. 08:30 < nmz787_i> kanzure: you didn't realize that was electrode-only? 08:30 < nottimschmidt> Keeps the realtime code small and fast. 08:31 < fenn> if you offload g-code to a uC you dont really need a pc at all 08:31 < nottimschmidt> fenn: added benefit 08:31 < nmz787_i> nottimschmidt: it's mostly the lag to come out of (kernel mode, I think) and go into (system, I think)... regardless of OS... just thinking of getting something from the CPU to the outside 08:32 < fenn> part of why people are resistant to using pc-based controllers in the first place i guess 08:32 < nmz787_i> it's a very x86 specific reason, at the bare-metal layer 08:32 < kanzure> nmz787_i: correct, i did not realize 08:32 < nottimschmidt> nmz787_i: there's nothing specific about x86 that causes this problem. 08:32 < nmz787_i> that's why the galileo/edison can only do a few 10s or 100s of kilohertz busy-loop GPIO toggling 08:32 < nottimschmidt> Attempting to do same on a big multi-core ARM would be just as bad. 08:32 < kanzure> fenn: linux everywhere is pretty nice.... 08:33 < nmz787_i> nottimschmidt: it definitely is, I've talked to experts 08:33 < fenn> nmz787_i: x86 io involves the ISA bus which has extremely slow timing for legacy reasons... 1ms round trip i think 08:33 < nottimschmidt> nmz787_i: experts, eh? 08:33 < nmz787_i> fenn: does a modern x86 even have ISA? 08:33 < fenn> there may be no actual ISA bus anywhere outside of the chip, but it's still used *shrug* 08:34 < fenn> this is what i was told several years ago and it's probably still true 08:34 < juri_> for some parallel ports. this is PCIE signaling. i think that problem is now dead. 08:34 < nottimschmidt> fenn: ISA doesn't exist any more in most x86 chips. The system management bus is logically compatible, but implemented on top of I2C 08:34 -!- Netsplit *.net <-> *.split quits: rigel, padz, dpk, mf1008, BobaMa, Betawolf, sh 08:34 < nottimschmidt> or SPI, forget. 08:35 -!- Netsplit *.net <-> *.split quits: heath, EnabrinTain, eleitl 08:35 -!- Netsplit over, joins: heath 08:35 -!- Netsplit over, joins: padz 08:35 < juri_> plus, linux has come up with some solutions to the X86 slowness too. kernel threads, for instance. 08:35 -!- Netsplit over, joins: Betawolf, eleitl, sh, mf1008 08:35 < juri_> <--- linux kernel hacker 08:36 -!- BobaMa [bobama@kapsi.fi] has joined ##hplusroadmap 08:36 -!- Netsplit over, joins: dpk 08:36 < fenn> juri_: any insight as to why linuxcnc still doesn't run on a beagleboard after all these years? 08:37 < nottimschmidt> fenn: I'm told there's some trouble with the beaglebone's GPIO implementation being done through sysfs 08:37 -!- Viper168 [~Viper@unaffiliated/viper168] has quit [Ping timeout: 264 seconds] 08:37 < nottimschmidt> It's possible to flip GPIOs without going through the FS layer, by peeking and poking various memory locations, but those change from kernel version to kernel version 08:38 < juri_> no good answer. i don't use it. it's based on the realtimelinux patches which disable ACPI (even on a realtime compliant COREBOOT system), so i'd start there... 08:38 < nottimschmidt> So far, no one's implemented a library to do it the fast way 08:39 < fenn> nottimschmidt: that's... not the answer i expected at all 08:40 < nmz787_i> CaptHindsight: I saw your quote, you listed solenoids... but I couldn't tell what for... what are they hooked up to? (Do I have to read through that POSAM paper to decipher the parts list?) 08:40 < CaptHindsight> juri_: http://openlunchbox.com/open-sbc/ imx6 with FPGA on the same board 08:40 < juri_> which FPGA? ;) 08:40 < nmz787_i> agreed --> 07:46 < ParahSailin> kanzure: and thats only because you emailed them before you emailed a cmos prototype shop 08:40 < CaptHindsight> nmz787: drying gas and reagents 08:41 < CaptHindsight> juri_: something that Mesa supports so that it can use all their open cores 08:42 < nmz787_i> CaptHindsight: please explain a bit more, are you using them as single-stroke piston pumps? valves? 08:42 < fenn> seems sort of backward to pick your open hardware to be able use existing semi-closed source software 08:42 < CaptHindsight> nmz787: it's the POSaM design 08:42 < nmz787_i> all reagents in the ABI 391 are under pressure, and valves open/close to let them around the machine 08:42 < juri_> CaptHindsight: I'm only interested in the xilinx spartan 6 slx9 or slx45. ;) 08:43 < fenn> juri plz pester the lunchbox people about this issue 08:43 < CaptHindsight> juri_: http://www.mesanet.com/fpgacardinfo.html 08:44 < juri_> yea, i have to judge from the jpegs, but it looks like they are using the right stuff. 08:44 < CaptHindsight> fenn: I'll use whatever hardware people pay for on the openlunchbox designs 08:44 < fenn> huh i thought it was supposed to have an fpga on board 08:44 < fenn> maybe i am mixing it up with some other open hardware laptop thingy 08:44 < fenn> novena? 08:45 < CaptHindsight> yeah, novena 08:45 -!- PatrickRobotham [uid18270@gateway/web/irccloud.com/x-duyxyyetgupmyrla] has quit [] 08:45 < fenn> The general purpose FPGA on the Novena mainboard is a Xilinx Spartan6 XC6SLX45 08:45 < nmz787_i> x86 will be soon seen with FPGAs on-chip 08:45 < chris_99> from intel? 08:45 < CaptHindsight> I offered to make the whole laptop but there isn't much interest outside a few paranoid or grey market folks 08:46 < fenn> really there isn't interest in an open laptop design? 08:46 < fenn> seems hard to believe 08:46 < kanzure> lkcl wants an open laptop, he dumped like $100k into some production runs 08:46 < CaptHindsight> actually had some people talk about evading law enforcement with the designs in IRC 08:47 < juri_> there's interest, but free hardware/software hackers are also cheap by necessity. 08:47 < fenn> if "evading law enforcement" means not having any factory-installed backdoors, i'm fine with that 08:47 < CaptHindsight> fenn: well it's up to AMD to reopen the AGESA firmware for the recent chips 08:47 < CaptHindsight> fenn: they were talking about smuggling 08:48 < fenn> smuggling what? 08:48 < fenn> actually nevermind, i don't care 08:48 < CaptHindsight> I tried to work with lkcl a few years ago, no thanks 08:49 < kanzure> nmz787_i: show me a single-nucleotide pump, or single-molecule pump from the literature, really. 08:49 < nmz787_i> chris_99: only speculation, but seems extremely likely 08:49 < kanzure> nmz787_i: i don't like you going around to out-of-band communication to try to convince me that you have this figured out 08:49 < chris_99> nice, i saw something about them supposed to make FPGA x86 chips a while ago iirc 08:49 < nmz787_i> kanzure: they do it all the time, dilute something to 1-molecule per milliliter... suck out 1 milliliter 08:49 < kanzure> nmz787_i: you can't handwave 08:49 < fenn> i would like an actually open laptop design, but i am cheap also 08:49 < kanzure> nmz787_i: he's not handwaving when he says "i'm using the posam valves and solenoid actuators". 08:50 < juri_> chris_99: there were FPGAs that fit into socket940. 08:50 < chris_99> http://www.extremetech.com/extreme/184828-intel-unveils-new-xeon-chip-with-integrated-fpga-touts-20x-performance-boost is what i was thinking of 08:50 < nmz787_i> kanzure: he didn't really reply though... I don't want to go read that paper when he already has the answer 08:50 < chris_99> juri this is an x86 with FPGA though 08:50 < juri_> nmz787_i: i read the paper yesterday. ;) 08:50 < nmz787_i> juri_: and I read it months and years ago 08:50 < juri_> it's a good read. i recommend it. 08:50 < fenn> still waiting for "reconfigurable computing" - the FPGA patent bog has delayed this miracle of technology for 20 years at least 08:51 < kanzure> CaptHindsight: hah, glad to hear you have met lkcl 08:51 < nmz787_i> juri_: meh, it's kind of boring to me outside the chemistry section 08:51 < juri_> lkcl follows my 3d printing work, but doesn't talk to me on irc, only email. 08:52 < juri_> weird. 08:52 < CaptHindsight> kanzure: have also met flyback, hours of fun 08:52 -!- EnabrinTain [sid11525@gateway/web/irccloud.com/x-vyhcaaiofprbcbch] has joined ##hplusroadmap 08:52 < juri_> flyback is troubled. 08:52 < kanzure> CaptHindsight: nmz787_i thinks that you haven't put enough thought into the valves and solenoids. 08:52 < juri_> I've known him for.. 15+ years. 08:52 < CaptHindsight> fenn: yeah, I've been hearing about it since the 80's 08:53 < fenn> but now we have "apps" 08:53 < fenn> woo 08:53 * fenn cries 08:53 < nmz787_i> kanzure: that isn't true... I simply asked if they were pumps or acting as valves 08:53 < kanzure> nmz787_i: gravity-assisted phosphoramidite dispense from the vials into the inkjet printhead 08:54 < nmz787_i> kanzure: so a siphon? 08:54 < CaptHindsight> nmz787: there's a manifold with solenoid vales, it's pretty simple 08:54 < nmz787_i> or this is still argon-pressure driven? 08:54 < CaptHindsight> it's like having multiple faucets going to one spout 08:55 < nmz787_i> that isn't an answer though to the question 08:55 < CaptHindsight> the paper jumps between argon and nitrogen, the point is no oxygen and no water 08:55 < juri_> I can talk to some people at BUGS, in case the 'where te put the machine' problem is still a problem. they have a wet lab there. 08:55 < CaptHindsight> nmz787: what you you really want to know? 08:55 < nmz787_i> you just answered it 08:55 < CaptHindsight> you/do 08:55 < nmz787_i> pressure-driven pumping, with the solenoids acting as valves 08:56 < juri_> a few ideas: 08:56 < juri_> you should be using the same laser for droplet detection as you use for head position checking. just make the mechanics interfere. 08:57 < CaptHindsight> electrically operated on/off vales 08:57 < juri_> the first priority should be droplet detection and analisys. 08:57 -!- PatrickRobotham_ [uid18270@gateway/web/irccloud.com/x-qfnipffmztvesdfv] has joined ##hplusroadmap 08:57 < kanzure> juri_: there's a droplet detector in the spec 08:57 < kanzure> specification 08:57 < CaptHindsight> juri_: we can also add a camera to look at drops 08:58 < juri_> we should be doing volumetric work as soon as we can verify we can fire the jets. so we can adapt the control signal to the fluids in question. 08:58 < nmz787_i> camera needed anyway for tritly color detection 08:58 < nmz787_i> trityl* 08:58 < CaptHindsight> and yes, the laser would tell you when the printhead is at a Home position 08:58 < fenn> lol yeah right "It is probably a lot easier to rewrite a small segment of critical code to run on an FPGA than to rewrite the entire application in OpenCL." 08:58 < juri_> ok, good. 08:58 -!- PatrickRobotham_ is now known as PatrickRobotham 08:58 < nmz787_i> unless you were underdosing the acid, and taking the efficiency hit for simplcitys sake 08:58 < fenn> openCL is super portable and has plenty of uses outside of GPGPU 08:58 < CaptHindsight> 2 lasers can give you XY 09:01 < CaptHindsight> laser are also often used to prevent the heads from crashing into the print substrates 09:02 < CaptHindsight> you don't want an idiot to crash a $8k printhead by not setting the lower limit of the Z 09:02 < CaptHindsight> or only checking one edge of the substrate when the other end is 5mm thicker 09:03 < juri_> it's a little different from what i do, where the worse damage is realigning, and scratches in the bed. 09:04 < CaptHindsight> we still don't want to smack a $200 printhead 09:04 < fenn> i want an all-out televised printhead brawl, with lasers and rockets 09:05 < CaptHindsight> it takes a few hours to get the printer back up again if you do it when you're ready to print with all the vials primed 09:06 < juri_> so, what do you have for signaling specs on these new heads? 09:07 < CaptHindsight> juri_: https://www.youtube.com/watch?v=hq-r0vAMURw watching drops in flight 09:08 < nmz787_i> fenn: battlebots -> printrbattle? or battleprintrs? 09:08 < kanzure> i'm still getting random email from the central bank of ecuador; they are still flustered that i asked for an sdk. 09:10 < kanzure> i need to decide about electrode array vs not electrode array. bleh, i don't want to have to manufacture increasingly smaller electrode arrays. that's going to be a pain in the butt. 09:11 < CaptHindsight> anyone know these people? https://www.youtube.com/watch?v=SloUgM6k77g BioDot BioJet Elite™: Drop-on-Dro 09:11 < kanzure> .title 09:11 < yoleaux> BioDot BioJet Elite™: Drop-on-Drop - YouTube 09:11 < nmz787_i> kanzure: fab is probably more accessible than those printheads, worldwide 09:11 < CaptHindsight> they use microvales for dispensing 09:12 < nmz787_i> well, at least in terms of more-than-one-off-pricing 09:13 < CaptHindsight> the heads may be removed from a new Epson 1430 $300 or for ~$200 from the Asian suppliers that assemble the printers 09:14 < CaptHindsight> there are a few importers that also sell just heads locally in the US and in the EU 09:15 < fenn> electrode array would be more appealing if it weren't exactly as slow as every other reaction chemistry out there 09:15 < fenn> can you even buy those chips? 09:15 < nmz787_i> no but the scales are huge in terms of fab 09:15 < CaptHindsight> there is a demand for just heads since people started hacking the Epson printers to make T-shirt printers 09:16 < kanzure> fenn: seem to be purchasable here https://azcobiotech.com/oligo-array-12k-chip-cleavable-19-5012.html 09:16 < nmz787_i> they'd be able to be produced on the oldest/cheapest process available at any place, most likely (outside academia, where they still like to train kids on 100+ micron features, just because it works and is easier) 09:17 < nmz787_i> CaptHindsight: do you think the plastic housing materials have changed since the POSAM paper? that would require re-tweaking of chemistry 09:17 < fenn> uh so is that reusable? 09:17 < kanzure> unclear if reusable 09:17 < kanzure> if you can cleave everything off of it.... 09:17 < CaptHindsight> DLP lithography can get us few micron features 09:17 < nmz787_i> not to mention every time the manufacturer decides to change materials, since that likely affects ink less than DNA synth chems 09:17 < fenn> .c 375/(12000*100) 09:18 < yoleaux> 375/(12000×100) = 1/3200 09:18 < fenn> bah 09:19 < fenn> well it's not a huge marginal cost even if it's not reusable, assuming you use the whole chip every time 09:19 < nmz787_i> kanzure: you realize you will have to do some damn costly sorting to get verbatim sequences out of an array right? you realize that was cambrian genomics whole ballgame, right? 09:20 < kanzure> sorting? 09:20 < fenn> still it would be annoying to have to wait until you have 12000*n bases to synthesize 09:20 < fenn> the point of having your own synthesizer is to shorten the development cycle right? 09:20 < nmz787_i> kanzure: well your 12k spots off 100bp would not be /only/ 100bp... it's going to be a spectrum of lengths 09:21 < nmz787_i> kanzure: you're still going to need to sort/filter the bad sequences, if you aren't doing some evolutionary work where some error is OK 09:21 < kanzure> fenn: interesting point 09:21 < kanzure> nmz787_i: that's the same case with cambrian's method anyway. it's just strands on beads. the beads can have many molecules that each have different errors. 09:22 < nmz787_i> well except that they were sequencing each bead, and only using the ones with low/no error 09:22 < nmz787_i> also, their beads may have been quite tiny, or primed in very dilute conditions so that few oligos grew per bead 09:23 < nmz787_i> kanzure: my point is... they weren't/aren't/haven't been so successful with their train of half-mil-$ machines 09:24 < kanzure> what? 09:24 < fenn> i really doubt the machine itself cost very much 09:24 < fenn> vs paying salaries for people for years, renting a space etc 09:25 < CaptHindsight> from what I saw of the printers they aren't that complex, not sure of that patent situation to make a DIY version 09:25 < nmz787_i> they had an azco machine, then a mi or hiseq, then their special Helicos-tech-like sequencer with laser desorption, probably an HPLC 09:28 < kanzure> fenn: if it's reusable then i think those concerns are nulled 09:29 -!- rigel [~yourmom@c-24-21-52-83.hsd1.or.comcast.net] has joined ##hplusroadmap 09:30 < CaptHindsight> oligo array 12K chip 50 pk Our price $16,250.00 09:30 < fenn> i'm unclear on how much reagents the array chip uses and how that compares to inkjet 09:30 < kanzure> i wonder why they would have a 50 pack 09:30 < kanzure> maybe they are reusable but people ship the arrays to other people for long-term use or something 09:31 < kanzure> (in which case, they should be rented to people, rather than just given) 09:31 < juri_> lead time to manufacture? 09:31 < fenn> you can use the arrays as hybridization detectors 09:31 < fenn> instead of cleaving them off 09:31 < kanzure> yes, i believe most people use these arrays for hybridization stuff 09:31 < fenn> say you want to detect if genes 1 to 12000 are present in a cell extract 09:35 < kanzure> ah they have a non-cleavable version? http://azcobiotech.com/oligo-array-12k-chip-non-cleavable-19-5013.html 09:35 < nottimschmidt> back, catching up 09:37 < kanzure> here are some weird ways the customarray microarray has been used by others http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3231185/ 09:38 < ParahSailin> nmz787_i: and the good part is, he doesnt even need to go to a cmos shop because it can be bought off the shelf as a disposable 09:38 < ParahSailin> heh, rice people bugging me again to pay off my parking tickets so that i can pick up my diploma after how many years? 09:39 < kanzure> here is church mentioning customarray http://arep.med.harvard.edu/pdf/Kosuri_Church_2014.pdf 09:39 < kanzure> "$0.00001/nucleotide" 09:41 < kanzure> "Array-based gene synthesis. Even though microarray-based oligo pools are cheap, there are several challenges in using them for gene synthesis. First, although the numbers of oligos that can be produced in a pool are large, their individual concentrations are quite low for most existing gene synthesis protocols. Second, the error rates for oligo pools are usually higher than those for column-synthesized oligos. Finally, the sheer number ... 09:42 < kanzure> ... of oligos produced leads to interference between gene assemblies, making it difficult to scale up." 09:42 < kanzure> "Tian et al.35 were the first to show how these problems could be overcome. They used PCR amplification to increase the concentration of the oligos before assembly, error-corrected them by hybridization to reverse complementary oligos (also constructed on the chip), and designed protein sequences computationally to avoid potential mishybridizations of the sequences." 09:42 < kanzure> "However, this work and contemporaneous reports36,37 used only dozens to hundreds of oligos at once. Scaling these methods proved difficult beyond pool sizes of 1,000 oligos38. At greater pool complexities, where the advantages in cost would come to play, constructing any individual gene became difficult, presumably owing to spurious cross-hybridization during the assembly process. In addition, the methods required sufficient sequence ... 09:42 < kanzure> ... orthogonality between synthesized genes, which limited potential applications." 09:42 < kanzure> "To alleviate these and other issues, two approaches were used that first isolated subpools of oligos required for any single assembly, thereby overcoming the concerns about both pool complexity and sequence orthogonality (Fig. 3). Kosuri et al.39 used predesigned barcodes that allowed PCR amplification of oligos participating in only a particular assembly and then removed the barcodes by digestions, which was followed by standard ... 09:42 < kanzure> ... assembly of the genes. Quan et al.40 used a custom inkjet synthesizer20 that synthesized subsets of oligos in physically separated microwells, where amplification and assembly were then done in situ. Both methods used much larger oligo pools (>10,000 oligos) and enzymatic error correction, which paved the way for commercialization in recent years (Gen9)." 09:43 < kanzure> well there's no reason to use a single pool 09:43 < fenn> the barcode thing is cool (and relevant to gene synthesis) 09:43 < kanzure> i don't know why people are focusing on single pool 09:44 < fenn> what's "enzymatic error correction"? 09:44 < ParahSailin> theres that dnase that attacks non-duplex dna 09:45 < kanzure> "More general methods of error correction usually depend upon a number of enzymatic techniques to reduce errors. All of these techniques rely on the fact that at any given position, most molecules possess the correct base. Heating and reannealing can force the formation of heteroduplexes that will contain disruptions to the canonical helical DNA structure. Such disruptions can be recognized and acted upon by several proteins." 09:45 < kanzure> "MutS binds heteroduplexes and can be used to filter errors by reverse purification13. Certain polymerases with exonuclease activity, endonucleases and resolvases can all cut or nick at such heteroduplex sites and, upon reamplification, can help filter errors12,26,45–47. Commercial enzymatic cocktails such as ErrASE have been commonly used to help reduce errors in synthetic genes as well30,39. Such error reduction can greatly reduce ... 09:45 < kanzure> ... the cost and time of gene synthesis by bringing error rates low enough that the genes can be directly used for functional assays without in vivo cloning and sequence verification30,48." 09:45 < fenn> ParahSailin: if you had a deletion mutation, wouldn't the dnase just destroy the opposing strand then? 09:46 < fenn> (not the mutated strand) 09:46 < kanzure> instead of single pool why isn't everyone using micropipette of each well? 09:46 < fenn> kanzure because it's a pain to synthesize things in wells 09:47 < kanzure> "More recent approaches have leveraged NGS technologies to screen and then select for perfect sequences at either the oligo or gene level. Matzas et al.49 used 454 sequencing (Roche) combined with a robotic pick-in-place pipette to mechanically pull reads with perfect sequence off the sequencing array and used these oligos for synthetic genes. Kim et al. also used 454 but marked individual molecules with random tags and then amplified ... 09:47 < kanzure> ... constructs with perfect sequence42. Both approaches help reduce error rates, although they still suffer from sequencing errors and require more expensive long-read platforms. Schwartz et al.41 used Illumina sequencing of barcodes to pull out perfect sequences but overcame limitations in length and sequencing errors through a tag-based consensus-sequencing approach. All three approaches substantially cut error rates and will continue ... 09:47 < kanzure> ... to improve with the rapid progress in DNA sequencing technologies. These NGS based error-correction approaches are also especially exciting for library-based constructions and synthesis methods because they allow correction without having to separate each gene assembly into individual reactions." 09:47 < kanzure> so.. that's wells/pores right there. 09:47 < kanzure> as far as i know 09:49 < kanzure> http://image.slidesharecdn.com/ngsintrov6public-110320062557-phpapp01/95/ngs-introv6public-15-728.jpg?cb=1300602396 09:49 < kanzure> (image is a cross-section of picotiter plates) 09:50 < kanzure> that's what we should be doing 09:50 < kanzure> this method is also compatible with inkjet stuff 09:50 < ParahSailin> thats ancient 09:50 < fenn> the pick and place thing is wells, the other two methods are not 09:50 < ParahSailin> do it like an illumina flow cell, its already attached to a glass slide 09:51 < fenn> i really dont get the microtiter thing 09:51 < ParahSailin> all you need then is lasers and microscopes 09:51 < kanzure> fenn: it's just a high-density pore array 09:51 < kanzure> and then they use pick-and-place 09:51 < fenn> they mechanically position a pipette tip to within 40 microns? 09:51 < kanzure> well, not for sequencing, but yes 09:52 < CaptHindsight> fenn: what do you see as the problems with wells? 09:52 < fenn> and then do that a zillion times? 09:52 < kanzure> fenn: that's my understanding of the last quote i pasted 09:52 < fenn> doesn't that seem ... crazy? 09:52 < kanzure> it's more specific than single pool 09:53 < juri_> CaptHindsight: is that an answer to my question? 09:53 < ParahSailin> what you really want is oligo clones 09:53 < CaptHindsight> heh yeah 09:53 < kanzure> matzas reference is "High-fidelity gene synthesis by retrieval of sequence-verified DNA identified using high-throughput pyrosequencing" 09:53 < ParahSailin> so load your pooled library into flowcell and do the bridge amplification step 09:53 < kanzure> http://arep.med.harvard.edu/pdf/Matzas_10.pdf 09:53 < ParahSailin> then sequence all the clusters on flow cell and pick and place 09:54 < fenn> CaptHindsight: have to add reagents to each well, and the wells get in the way of your deposition method (?) 09:55 < ParahSailin> destroy all bad clusters with uv laser to make it easier to pick good ones off 09:55 < fenn> there's more area available for synthesis if there are no well walls 09:55 < fenn> i'm not really claiming these are obstacles 09:56 < ParahSailin> no, that probably wouldnt work 09:56 < kanzure> i agree that pore/well walls are not entirely necessarily 09:56 < nmz787_i> kanzure: micropipette of each well would still be a single pool though 09:56 < kanzure> especially if you are doing single drop isolation 09:56 < kanzure> nmz787_i: what?? 09:56 < CaptHindsight> fenn: ok the walls take up space, so wall-less wells use surface tension 09:56 < ParahSailin> illumina has gone to patterned flow cells with wells 09:56 < nmz787_i> you can have a kiddie pool or an olympic sized pool, both are pools 09:56 < kanzure> nmz787_i: yes, each drop is a single pool, so what? 09:57 < fenn> CaptHindsight: i hadnt considered that 09:57 < nmz787_i> I'm guessing their assumptions hold for all sized pools, minus the pool where there are few enough molecules to count them on your fingers and maybe toes also 09:57 < kanzure> ParahSailin: can you please find me a useful diagram of an illumina flow cell :-( i don't understand 09:58 < ParahSailin> http://www.illumina.com/company/video-hub/pfZp5Vgsbw0.html 09:58 < kanzure> .title 09:58 < yoleaux> Patterned Flow Cell Technology | Illumina Video 09:58 < juri_> CaptHindsight: so, is that answer "i've got it"? 09:58 < fenn> .title http://youtu.be/pfZp5Vgsbw0 09:58 < yoleaux> Patterned Flow Cell Technology | Illumina Video - YouTube 10:00 < ParahSailin> you can get scrap used flow cells in the trash 10:00 < nmz787_i> kanzure: you were complaining about people talking about single pools... I am guessing they are concerned with single pools because they need this info to be able to say whether or not the process can be scaled to multiple parallel pools, or sequential, etc 10:00 < fenn> ooo aah 10:01 < kanzure> nmz787_i: the point is that you don't want all dna from all pores in the same pot or pool. you can't assemble genes easily in that scenario. but if you separately handle each pore, you can combine strands together at your own pace. 10:01 < fenn> they ensure only one sequence per well simply by reaction kinetics eh? 10:01 < kanzure> nmz787_i: no they are doing typical dna hybridization stuff.... they aren't doing gene assembly. 10:01 < kanzure> fenn: yes that sort of surprises me 10:02 < fenn> no it makes sense 10:02 < fenn> alright i am ready to unlock the power of the genome 10:03 < ParahSailin> i dont know what fraction of the wells randomly start with one oligo 10:03 < archels> "Hi all, I’m writing to inform you all that in the cell culture lab I dark reared some mice in the drawer where the syringes are stored. Please do not open or attempt to open the above mentioned drawer." 10:03 < ParahSailin> thats more of a matter of tinkering with the library dna concentration 10:03 < fenn> "a high percentage" 10:04 < fenn> there was some kind of initiator molecule, so if you have a low concentration of that, and a low concentration of dna, it's extremely unlikely that you'd have two dna's and two initiators at the same place and time 10:04 < kanzure> "with enough scale the error rate doesn't matter!~~~" 10:04 < fenn> once there's one initiator and one dna, amplification happens quickly and the substrate gets used up 10:04 < nmz787_i> fenn: likely some non-polymer, so they can ensure a monolayer? 10:05 < fenn> nmz787_i: what? 10:05 < nmz787_i> the initiator 10:05 < fenn> oh, it looked like an enzyme 10:05 < fenn> it was a red blob 10:05 < ParahSailin> guys illumina publishes protocols 10:06 < fenn> all i know is what we just saw http://youtu.be/pfZp5Vgsbw0#t=54s 10:06 < ParahSailin> http://support.illumina.com/content/dam/illumina-support/documents/myillumina/ab41723c-129f-47e1-8000-7282fb605c87/clusterstationuserguide_15018818_d.pdf 10:07 < nmz787_i> .youtube gcta pcr commercia 10:07 < kanzure> pick-and-place bead protocol could be done on the same microarray plate. you could probably run lots of simultaneous gibson assemblies or something. 10:07 -!- nmz787_i [ntmccork@nat/intel/x-nxxygqsseduszlkg] has quit [Quit: Leaving.] 10:08 < CaptHindsight> exclusion amplification 10:08 < CaptHindsight> what is this magic? 10:10 < CaptHindsight> http://www.google.com/patents/WO2013188582A1?cl=en 10:10 < CaptHindsight> Kinetic exclusion amplification of nucleic acid libraries 10:11 < fenn> i wonder why this took until 2013 to figure out 10:11 < fenn> pretty sure it is older than that 10:11 < ParahSailin> illumina has been commercial since 2008 10:12 < CaptHindsight> maybe they just decided to patent after the lawyers combed through everything 10:13 < fenn> illumina is a company; are you saying they only use this specific sequencing method based on exclusion amplification? 10:13 < ParahSailin> oh, its not clear what this kinetic exclusion bla is 10:13 < ParahSailin> clearly not essential to flowcell sequencing 10:14 < kanzure> all the cells are under the same flow? 10:14 < kanzure> so they are experiencing aqueous contamination with each other? 10:14 < ParahSailin> there are only 8 different channels in a flow cell 10:17 < kanzure> so... yes? 10:19 < ParahSailin> each cluster on the flow cell is in the same aqueous solution 10:20 < kanzure> fenn: here is the "crazy" micropippete + microarray pick-and-place approach, see figure 2 on page 2 http://www.nature.com/nbt/journal/v28/n12/extref/nbt.1710-S1.pdf 10:21 < kanzure> oh is that the same matzas paper. i see. 10:21 < CaptHindsight> would it help to have a femtoliter drop size vs pL? 10:21 < fenn> that's a really thick block of granite 10:21 < fenn> don't sneeze 10:22 < CaptHindsight> heh 10:23 < fenn> so this actually works in practice? not just like one or two sequences 10:23 < kanzure> i seem to recall that they had a person operating the pipette tip, and they were manually looking at the video to determine when they were crashing the tip into the edge of a well or pore 10:24 < kanzure> but i don't remember why they were doing this 10:24 < fenn> i gotta say i like the pcr barcode way just in principle 10:24 < kanzure> because at 4096x4096 that's.. a lot of video game playing. 10:25 < fenn> from DNA select * where sequence=^AGCT.* 10:26 < CaptHindsight> fenn: here's a 5-axis we did for a large silicon valley fruit company http://imagebin.ca/v/28Vp4S0dYt00 10:26 < kanzure> is this just dna hybridization wings technique 10:26 < kanzure> that should be labeled nsfw, it's clearly machine porn 10:26 < CaptHindsight> 5um repeatability with inkjet, lasers and microscope 10:27 < fenn> i can almost smell the ethylene 10:28 < fenn> ok i think my brain is melting bbl 10:29 < CaptHindsight> we can pick and place using a 0.5um tip pretty quick using linear servos 10:30 < kanzure> i wonder why they needed manual operator control of that 10:30 < CaptHindsight> budget, skill ? 10:31 < CaptHindsight> time 10:33 < CaptHindsight> heh pages 23- 135 are all just the sequences in text format 10:33 < kanzure> yes, biologists are silly 10:33 < CaptHindsight> I winder if that would fit on a floppy? 10:35 < kanzure> "Kosuri et al.39 used predesigned barcodes that allowed PCR amplification of oligos participating in only a particular assembly and then removed the barcodes by digestions, which was followed by standard assembly of the genes." 10:35 < kanzure> "Quan et al.40 used a custom inkjet synthesizer20 that synthesized subsets of oligos in physically separated microwells, where amplification and assembly were then done in situ." 10:35 < CaptHindsight> maybe inkjet directly to beads 10:35 < kanzure> yes 10:35 < kanzure> totally doable 10:35 < kanzure> ( those quotes are from http://arep.med.harvard.edu/pdf/Kosuri_Church_2014.pdf ) 10:36 < CaptHindsight> beads + laser launcher 10:36 < CaptHindsight> lets get the chems worked out and the timing 10:36 < CaptHindsight> then we can print on beads 10:37 < CaptHindsight> how did Cambrian make their beads? 10:38 -!- nottimschmidt [~timschmid@35.10.220.177] has quit [Ping timeout: 265 seconds] 10:40 < juri_> bbiaf. 10:42 < kanzure> CaptHindsight: there's lots of places to buy beads 10:45 -!- nmz787_i [~ntmccork@192.55.54.42] has joined ##hplusroadmap 10:49 -!- nmz787_i1 [~ntmccork@134.134.139.72] has joined ##hplusroadmap 10:50 -!- nmz787_i [~ntmccork@192.55.54.42] has quit [Ping timeout: 255 seconds] 10:51 < CaptHindsight> hmm, printing right onto beads might only help if you have very low error rates 10:53 -!- nottimschmidt [~timschmid@13-195-195.client.wireless.msu.edu] has joined ##hplusroadmap 10:54 < kanzure> the beads is more for easy transport 10:54 < kanzure> afaik it does not materially impact things like reaction kinetics 10:54 < CaptHindsight> a laser can launch them off anything 10:56 < kanzure> btw you might be unaware but there are some pcr techniques that involve laser heating of picoliter droplets on a surface 10:57 < CaptHindsight> makes sense 10:57 < nmz787_i1> well beads can have a multitude of surface material modifications 10:58 < nmz787_i1> CaptHindsight: you can take a look at bangslabs.com 10:59 < kanzure> "Parallel on-chip gene synthesis and application to 10:59 < kanzure> gah 11:00 < kanzure> "Parallel on-chip gene synthesis and application to optimization of protein expression" http://www.researchgate.net/profile/Nicolas_Negre/publication/51073109_Parallel_on-chip_gene_synthesis_and_application_to_optimization_of_protein_expression/links/0046352567745e4375000000.pdf 11:00 < kanzure> they are claiming oligo amplification and assembly on the same inkjet-printed array? 11:00 -!- delinquentme [~delinquen@74.61.157.78] has joined ##hplusroadmap 11:02 < kanzure> ah, polymerase cycling assembly 11:03 < kanzure> er, "polymerase chain assembly" 11:10 < kanzure> .title http://www.nature.com/ncomms/2015/150202/ncomms7073/full/ncomms7073.html 11:10 < yoleaux> A high-throughput optomechanical retrieval method for sequence-verified clonal DNA from the NGS platform : Nature Communications : Nature Publishing Group 11:10 < kanzure> "pulse laser retrieval system" 11:10 < kanzure> "Writing DNA plays a significant role in the fields of synthetic biology, functional genomics and bioengineering. DNA clones on next-generation sequencing (NGS) platforms have the potential to be a rich and cost-effective source of sequence-verified DNAs as a precursor for DNA writing. However, it is still very challenging to retrieve target clonal DNA from high-density NGS platforms. Here we propose an enabling technology called ... 11:10 < kanzure> ... ‘Sniper Cloning’ that enables the precise mapping of target clone features on NGS platforms and non-contact rapid retrieval of targets for the full utilization of DNA clones. By merging the three cutting-edge technologies of NGS, DNA microarray and our pulse laser retrieval system, Sniper Cloning is a week-long process that produces 5,188 error-free synthetic DNAs in a single run of NGS with a single microarray DNA pool. We ... 11:10 < kanzure> ... believe that this technology has potential as a universal tool for DNA writing in biological sciences." 11:11 < kanzure> a more general article, http://www.biotechniques.com/news/biotechniquesNews/biotechniques-356987.html#.VaP_rlIX3RY 11:11 < CaptHindsight> ooh 11:11 < kanzure> oh this seems to be the same person (bang) 11:11 < kanzure> nope nevermind 11:12 < kanzure> "Fortunately, they came across a 2010 Nature Biotechnology paper (1) from George Church’s group describing megacloning, which uses NGS to directly select sequence-verified clones from a complex pool of mixed oligonucleotides after NGS without the need for subsequent selective PCR amplification. This was possible using Roche 454 Life Sciences’ GLS platform, where each individual bead in a well in the open-top 454-Picotiterplate (PTP) ... 11:12 < kanzure> ... that corresponds to a sequenced oligonucleotide clone could be directly physically extracted with a micropipette using a robotic imaging and bead picking system." 11:12 < kanzure> "“The concept of direct utilization of clones on the NGS substrate was a refreshing jolt,” said Kwon, but as an engineer, he felt that there was room for improvement to the method. The throughput was constrained by the need for physical contact-based retrieval of the beads, making the method inadequate for generating the large number of oligonucleotides necessary for writing megabase-sized DNA pieces." 11:12 -!- ebowden [~ebowden@101.177.244.210] has joined ##hplusroadmap 11:13 < kanzure> "Kwon and Bang’s improved version of the megacloning method, Sniper Cloning, was recently published in Nature Communications (2). Their key modification is the use of a fully automated pulse laser system for the non-contact and very rapid retrieval of the desired beads from the PTP. They cleverly took advantage of the structure of the PTP, where each well that holds one bead is formed by etching of the end of an individual fiber optic ... 11:13 < kanzure> ... core that conducts the light released from the bead during pyrosequencing back to the CCD camera. By inverting the PTP, a low-energy laser pulse targeted to the back side (i.e. top) of a selected well is transmitted by the fiber optic core to the bead, pushing it out by radiation pressure down into the well of a 96-well microtiter plate positioned underneath the PTP. This approach greatly reduces the possibility of ... 11:13 < kanzure> ... cross-contamination inherent to megacloning and, by using an automated motorized stage, is orders of magnitude faster than retrieving the selected beads from the PTP." 11:14 < CaptHindsight> laser prod 11:14 < CaptHindsight> non-contact 11:14 < CaptHindsight> except for the well material 11:14 < kanzure> hmm so what doesn't make sense to me here is that the 96 array doesn't allow you much room... sure, you can punch any bead you want into any one of the 96 slots, then you can run further chemistry to combine the oligos. but then what if you want to combine two of those sequences together (to make a larger dna fragment)? 11:16 < CaptHindsight> it's 96 beads since the beads are large 11:16 < kanzure> they launch the smaller beads into the larger 96 plate 11:17 < CaptHindsight> inkjet pools to laser lanes 11:17 < kanzure> right, still need the inkjet to print on to the tiny beads anyway 11:18 < CaptHindsight> can't you just push the next oligo onto the last one combined to make it longer? 11:18 < CaptHindsight> what makes it stop at only 2? anything? 11:19 < kanzure> 96 plate is incompatible with laser transportation of beads into other 96 plate holes 11:20 < kanzure> there are 2 plates. super-tiny-pore plate is on top. 96 plate is below. stage moves 96 plate. laser pokes a micro bead out of tiny plate, drops into known pore on 96 plate. 11:20 < kanzure> you oculd use pipettes to transport material from one 96 plate pore to another pore on the 96 plate, i guess 11:24 < delinquentme> If the constant region is replaced with the human form, the antibody is termed chimeric and the substem used is -xi-. 11:24 < yoleaux> 10 Jul 2015 12:20Z delinquentme: https://www.youtube.com/watch?v=JiyxPae4h5A&t=17m30s 11:24 < delinquentme> do we know anyone who knows this to be decidedly true? ( its from wikipedia ) 11:24 -!- gnusha [~gnusha@unaffiliated/kanzure/bot/gnusha] has quit [Ping timeout: 255 seconds] 11:24 -!- gnusha [~gnusha@unaffiliated/kanzure/bot/gnusha] has joined ##hplusroadmap 11:24 -!- Topic for ##hplusroadmap: biohacking, nootropics, transhumanism, open hardware | sponsored by lobsters everywhere, banned by the Federal Death Administration (5 times) | this channel is LOGGED: http://gnusha.org/logs | http://diyhpl.us/wiki | "ray kurzweil is a pessimist" - george church 11:24 -!- Topic set by kanzure [~kanzure@unaffiliated/kanzure] [Wed May 20 12:46:25 2015] 11:24 [Users ##hplusroadmap] 11:24 [ abetusk ] [ catern ] [ EnLilaSko ] [ marchtemp ] [ poohbear ] [ superkuh ] 11:24 [ Acty ] [ chris_99 ] [ eudoxia ] [ mf1008 ] [ Qfwfq ] [ Taek ] 11:24 [ Adlai ] [ crescendo ] [ fenn ] [ midnightmagic ] [ redlegion ] [ the8thbit] 11:24 [ altersid ] [ Daeken ] [ gnusha ] [ nickjohnson ] [ rigel ] [ ThomasEgi] 11:24 [ andytoshi ] [ delinquentme] [ Guest34971 ] [ night ] [ ryankarason ] [ thundara ] 11:24 [ archels ] [ diginet ] [ heath ] [ nmz787 ] [ saurik ] [ TMA ] 11:24 [ augur ] [ dingo_ ] [ helleshin ] [ nmz787_i1 ] [ sh ] [ Urchin ] 11:24 [ Bakkot ] [ Douhet ] [ HEx1 ] [ nottimschmidt ] [ sheena ] [ vivi ] 11:24 [ balrog ] [ dpk ] [ JayDugger ] [ nsh ] [ sivoais ] [ wrldpc1 ] 11:24 [ berndj ] [ drethelin ] [ jrayhawk ] [ p4nd4 ] [ smeaaagle ] [ xrr ] 11:24 [ Betawolf ] [ drewbot_ ] [ juri_ ] [ padz ] [ strages ] [ yoleaux ] 11:24 [ bkero ] [ dustinm ] [ justanotheruser] [ panax ] [ strangewarp_] [ yorick ] 11:24 [ BobaMa ] [ ebowden ] [ juul ] [ ParahSailin ] [ streety ] 11:24 [ Burn_ ] [ eleitl ] [ kanzure ] [ pasky ] [ Stskeeps ] 11:24 [ CaptHindsight] [ EnabrinTain ] [ kish ] [ PatrickRobotham] [ super` ] 11:24 -!- Irssi: ##hplusroadmap: Total of 87 nicks [0 ops, 0 halfops, 0 voices, 87 normal] 11:24 -!- Channel ##hplusroadmap created Thu Feb 25 23:40:30 2010 11:24 -!- Irssi: Join to ##hplusroadmap was synced in 12 secs 11:24 -!- heath [~heath@unaffiliated/ybit] has quit [Ping timeout: 255 seconds] 11:26 -!- heath [~heath@unaffiliated/ybit] has joined ##hplusroadmap 11:26 < nmz787_i1> kanzure: it would be foreseeable to swap the electrochemical acid generation electrodes with some sensor-frontend and elucidate some data to increase synthesis fidelity 11:27 < kanzure> swap? 11:28 < nmz787_i1> from push to sense 11:28 < nmz787_i1> pin mux 11:28 < kanzure> ok pin mux 11:30 -!- nottimschmidt [~timschmid@13-195-195.client.wireless.msu.edu] has quit [Ping timeout: 264 seconds] 11:36 < kanzure> stalk: bernellev@gmail.com 11:38 -!- CheckDavid [uid14990@gateway/web/irccloud.com/x-egeeezwmfnjhahee] has joined ##hplusroadmap 11:47 -!- nottimschmidt [~timschmid@13-74-117.client.wireless.msu.edu] has joined ##hplusroadmap 11:47 -!- ant4t [~s3an@50.141.78.33] has joined ##hplusroadmap 11:55 -!- balrog [~balrog@unaffiliated/balrog] has quit [Ping timeout: 240 seconds] 12:00 < nmz787_i1> jrayhawk: are you or other paleo-types visually artistic at all? I am thinking it would be fun to have a shirt that reads 'I pass on grass' and has pictures of grain at the top of a stalk, popcorn, bread, corn syrup 12:01 < nmz787_i1> also rice 12:02 < nmz787_i1> maybe a picture of someone 'passing' it off to a cow 12:02 < nmz787_i1> 'pass the grass to the left-hand cow' 12:03 < CaptHindsight> from the Sniper paper "To the best of our knowledge, it is very difficult to supply a massive amount of synthetic oligonucleotides of extremely high standard, except through conventional cloning and random pick-and-place followed by individual identification" 12:04 < kanzure> random? 12:04 < nmz787_i1> i guess there are too many in their opinion to search each and every one 12:05 < nmz787_i1> and there's no better heuristic than random? 12:05 < CaptHindsight> heh, yeah random, just let the machine run wild 12:05 -!- nottimschmidt [~timschmid@13-74-117.client.wireless.msu.edu] has quit [Ping timeout: 240 seconds] 12:05 < kanzure> ok sounds like they mean "other than doing the full entire array, just use some random subset, because we don't want to wait for pick-and-place" 12:05 < kanzure> machines can pick-and-place very very quickly 12:06 -!- balrog [~balrog@unaffiliated/balrog] has joined ##hplusroadmap 12:07 < CaptHindsight> https://www.youtube.com/watch?v=u6KW8fIBjr8 Adept delta bot 12:08 -!- PatrickRobotham [uid18270@gateway/web/irccloud.com/x-qfnipffmztvesdfv] has quit [Quit: Connection closed for inactivity] 12:08 < CaptHindsight> https://www.youtube.com/watch?v=0-Kpv-ZOcKY this is the one from a few years ago handling ball bearings 12:21 < kanzure> "It is stinky shit, but it's kinda working." - eleitl 12:22 -!- nottimschmidt [~timschmid@13-65-55.client.wireless.msu.edu] has joined ##hplusroadmap 12:27 -!- davm23 [~davm@adsl-107-203-26-177.dsl.tul2ok.sbcglobal.net] has joined ##hplusroadmap 12:32 < CaptHindsight> with laser interferometry, linear servos and temp control we can have few um repeatability at high speeds with a femtoliter inkjet 12:33 < CaptHindsight> lets just say 5um between centers for sample spots 12:33 < nmz787_i1> at that point we'd get further by just making a cheap photolithography mastering system 12:33 < nmz787_i1> using a bluray drive optical sled 12:33 < nmz787_i1> (they have near-diffraction limit optics) 12:34 < nmz787_i1> http://diyhpl.us/~bryan/papers2/optics/photolithography/High%20resolution,%20low%20cost%20laser%20lithography%20using%20a%20Blu-ray%20optical%20head%20assembly.pdf 12:34 < CaptHindsight> doesn't lithography mean flooding each layer at a time? 12:34 < CaptHindsight> and having to move masks around? 12:35 < nmz787_i1> not necessarily... it can mean making microfluidics in a closed-system 12:35 < CaptHindsight> oh, yeah different animal 12:35 < CaptHindsight> easy once i make the machines to make those machines 12:36 < nmz787_i1> lot more microfluidics research than just DNA (i.e. cell culturing) 12:37 < CaptHindsight> the blue ray sleds are nice for the laser diode but they rely on tracks for accuracy 12:37 < nmz787_i1> nah that's only for positioning 12:37 < nmz787_i1> the focus is sub-micron 12:37 < nmz787_i1> that's why i commented on that only after you mentioned interferometry 12:37 < CaptHindsight> well depends on how we define the sled 12:37 < nmz787_i1> the part with the optics, that slides 12:37 < CaptHindsight> sure, we use them all the time 12:38 < CaptHindsight> the 405nm laser diodes 12:39 < nmz787_i1> well the diodes themselves need quite a bit of work to get to sub-micron focus 12:39 < CaptHindsight> why hasn't anyone made a low cost diy blueray printer for sub-micron features? 12:39 < nmz787_i1> TEM00 and such 12:39 < nmz787_i1> CaptHindsight: because time on my part 12:39 < CaptHindsight> do I have to do everything? 12:40 < nmz787_i1> my idea without interferometry was to use a grid reticle under a video microscope, and machine vision... for feedback 12:40 < nmz787_i1> and something like a microscope stage for XY 12:42 < CaptHindsight> how many would use it or build it? 12:43 < CaptHindsight> my concern is that you give the plans out and people can't make it work since they have too little experience with fabrication anything 12:44 < kanzure> CaptHindsight: yes a future machine worth making is a way to easily and quickly make microfluidic devices. either laser diode laser cutter or maskless photolithography or something. 12:44 < CaptHindsight> then you have to spend lots of time holding their hands to make it work 12:44 < kanzure> CaptHindsight: we had a partial laser diode cutter thingy, but we couldn't do the optics 12:44 < kanzure> http://diyhpl.us/laser_etcher/laser_etcher/ 12:45 < CaptHindsight> maybe give them the working mechanism and then they can wrap it and write software 12:46 -!- proofoflogic [uid65184@gateway/web/irccloud.com/x-wqcoozsjjgnawftq] has joined ##hplusroadmap 12:50 -!- MrHindsight [~not_sure@unaffiliated/capthindsight] has joined ##hplusroadmap 12:56 -!- genehacker [~chatzilla@128-193-154-1.ptpg.oregonstate.edu] has joined ##hplusroadmap 13:08 < kanzure> "Automatic error elimination by horizontal code transfer across multiple applications" http://people.csail.mit.edu/fanl/papers/codephage-pldi2015.pdf 13:16 -!- helleshin [~talinck@66-161-138-110.ubr1.dyn.lebanon-oh.fuse.net] has quit [Read error: Connection reset by peer] 13:19 -!- nmz787_i1 [~ntmccork@134.134.139.72] has quit [Quit: Leaving.] 13:21 -!- saurik [saurik@carrier.saurik.com] has quit [Ping timeout: 255 seconds] 13:21 -!- helleshin [~talinck@66-161-138-110.ubr1.dyn.lebanon-oh.fuse.net] has joined ##hplusroadmap 13:21 -!- saurik [saurik@carrier.saurik.com] has joined ##hplusroadmap 13:26 -!- nottimschmidt [~timschmid@13-65-55.client.wireless.msu.edu] has quit [Ping timeout: 256 seconds] 13:28 -!- Beatzebub [~beatzebub@d162-156-106-225.bchsia.telus.net] has joined ##hplusroadmap 13:30 -!- nmz787_i [ntmccork@nat/intel/x-fmxyjytcpyrvqtsx] has joined ##hplusroadmap 13:31 -!- Acty [uid89656@gateway/web/irccloud.com/x-lnomihmgkbtswyha] has quit [Quit: Connection closed for inactivity] 13:31 < nmz787_i> kanzure: we didn't stop the laser etcher because of the optics, did we? I thought fenn dropped offline and we didn't have anyone to help with CNC advice and modelling (and now fenn is less interested) 13:31 < nmz787_i> CaptHindsight: yes I think retrofitting a microscope is easy enough for the keen biologists 13:32 < nmz787_i> orders of magnitude moreso than a POSAM or derivative 13:32 < kanzure> no clue, i suppose that could be true? hm... 13:33 < nmz787_i> CaptHindsight: the microscope idea has been partially DIYed... http://openlabtools.eng.cam.ac.uk/Instruments/Microscope/ if sourcing microscopes to retrofit doesn't sound as appealing and repeatable as making your own 13:34 < nmz787_i> huh http://www.meetup.com/Cambridge-Synthetic-Biology-Meetup/events/221680133/ 13:34 < nmz787_i> .title 13:34 < yoleaux> Hack the Lab: how to build a microscope - Cambridge Synthetic Biology Meetup (Cambridge, England) - Meetup 13:35 < nmz787_i> .title http://www.meetup.com/Makespace/events/221524812/ 13:35 < yoleaux> Hack the Lab: how to build a microscope - Makespace (Cambridge, UK) (Cambridge, England) - Meetup 13:38 < kanzure> fenn: it just occurred to me that the pcr barcode method is not the same thing as dna hybridization overlap wings, the barcode is probably meant to be optimally-chosen for being different from all the other sequences, isn't it.... whereas overlap hybridization is just sharing relevant sequence between multiple strands. 13:40 -!- CheckDavid [uid14990@gateway/web/irccloud.com/x-egeeezwmfnjhahee] has quit [Quit: Connection closed for inactivity] 13:43 -!- nmz787_i1 [ntmccork@nat/intel/x-yjgbgkbahcqbepnl] has joined ##hplusroadmap 13:46 -!- nmz787_i [ntmccork@nat/intel/x-fmxyjytcpyrvqtsx] has quit [Ping timeout: 252 seconds] 14:11 -!- Acty [uid89656@gateway/web/irccloud.com/x-sozxtwlywxurstxe] has joined ##hplusroadmap 14:15 -!- Madplatypus [uid19957@gateway/web/irccloud.com/x-ktdebwyjroxkwhns] has joined ##hplusroadmap 14:15 -!- nmz787_i [~ntmccork@134.134.139.72] has joined ##hplusroadmap 14:16 -!- nmz787_i1 [ntmccork@nat/intel/x-yjgbgkbahcqbepnl] has quit [Ping timeout: 248 seconds] 14:20 -!- genehacker [~chatzilla@128-193-154-1.ptpg.oregonstate.edu] has quit [Ping timeout: 248 seconds] 14:24 -!- justanotheruser [~Justan@unaffiliated/justanotheruser] has quit [Ping timeout: 246 seconds] 14:25 -!- nottimschmidt [~timschmid@13-181-165.client.wireless.msu.edu] has joined ##hplusroadmap 14:37 -!- nmz787_i1 [ntmccork@nat/intel/x-qnyydaamforoisgn] has joined ##hplusroadmap 14:39 -!- nmz787_i [~ntmccork@134.134.139.72] has quit [Read error: Connection reset by peer] 14:41 -!- nmz787_i [~ntmccork@134.134.139.72] has joined ##hplusroadmap 14:43 -!- nmz787_i1 [ntmccork@nat/intel/x-qnyydaamforoisgn] has quit [Ping timeout: 256 seconds] 14:55 -!- genehacker [~chatzilla@128-193-152-178.ptpg.oregonstate.edu] has joined ##hplusroadmap 14:59 -!- chris_99 [~chris_99@unaffiliated/chris-99/x-3062929] has quit [Remote host closed the connection] 14:59 < nmz787_i> if there is an API call I'd like to block in a Windows executable, how would I skip it using a hexeditor? replace the call with a NOP or something? 15:03 -!- nottimschmidt [~timschmid@13-181-165.client.wireless.msu.edu] has quit [Ping timeout: 256 seconds] 15:07 -!- eudoxia [~eudoxia@r167-56-36-161.dialup.adsl.anteldata.net.uy] has quit [Quit: Leaving] 15:09 < juri_> nmz787_i: on the laser/CNC front, remember that i've been here to help with CNC and modeling for some time. just building up skills, waiting for this place to need me. 15:13 < juri_> I'm still no biologist, but when it comes to plastic printers, i know a thing or five hundred. 15:18 < nmz787_i> juri_: any noticeable improvement with the CAD repo? I remember the last time I tried it, it was giving pretty poor quality on the small features I was designing, and would have random 'holes' in the mesh 15:19 < juri_> nothing very notable. I'm still polishing the code, to make it easy enough for me to make radical changes. 15:20 < juri_> learning haskell, while fun, has been grueling. 15:21 -!- Viper168 [~Viper@unaffiliated/viper168] has joined ##hplusroadmap 15:23 < kanzure> nmz787_i: yes, you can insert nops 15:24 < kanzure> also, use idapro 15:34 -!- MrHindsight [~not_sure@unaffiliated/capthindsight] has quit [Quit: Leaving] 15:36 -!- nottimschmidt [~timschmid@2601:405:4101:52db::822] has joined ##hplusroadmap 15:37 -!- Darius [~quassel@75.76.19.18] has joined ##hplusroadmap 15:45 -!- genehacker [~chatzilla@128-193-152-178.ptpg.oregonstate.edu] has quit [Ping timeout: 256 seconds] 15:59 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has joined ##hplusroadmap 16:12 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has quit [Ping timeout: 250 seconds] 16:15 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has joined ##hplusroadmap 16:15 -!- ant4t [~s3an@50.141.78.33] has quit [Ping timeout: 264 seconds] 16:24 < nmz787_i> kanzure: my nature-based calendar tells me a plover is a bird that hangs out in the NYC area 16:24 < nmz787_i> .wik plover 16:24 < yoleaux> "Plovers (/ˈplʌvər/ or /ˈploʊvər/) are a widely distributed group of wading birds belonging to the subfamily Charadriinae." — https://en.wikipedia.org/wiki/Plover 16:28 -!- ant4t [~s3an@50.141.77.140] has joined ##hplusroadmap 16:34 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has quit [Ping timeout: 250 seconds] 16:56 < kanzure> nmz787_i: yes, it's a bird. the plover chording system was named after the bird. 16:56 < kanzure> nmz787_i: why can't i find any evidence of single-molecule dilutions + pumping in the literature? 16:56 -!- ant4t [~s3an@50.141.77.140] has quit [Ping timeout: 256 seconds] 17:01 < kanzure> isn't this some sort of brownian motion thermodynamics limit 17:01 < kanzure> andytoshi: aren't you a physics person 17:04 -!- erasmus [~esb@unaffiliated/erasmus] has joined ##hplusroadmap 17:11 < nmz787_i> kanzure: the dilution thing is pretty common 17:11 < nmz787_i> that's how they dilute hybridomas for making monoclonal antibodies 17:12 < nmz787_i> s/dilute/isolate/ 17:12 < CaptHindsight> we can start with an aluminum oxide filter with ~20nm pores 17:12 < nmz787_i> CaptHindsight: for? 17:12 < CaptHindsight> push the base through, and catch them a few at a time 17:13 < CaptHindsight> then we send that through a second pore and isolate the single molecules 17:14 < nmz787_i> wouldn't that just act to slow the flow down though? they aren't that much larger than a solvent molecule 17:15 < nmz787_i> hrmm, I've only ever really thought about it in terms of dilution and watching for it to pass by 17:15 < nmz787_i> aside from the molecular pump stuff, which I've stayed away from thinking much about 17:15 < CaptHindsight> well dilution helps to reduce the number of the molecules you want to isolate 17:17 < kanzure> isn't a single molecule pump supposed to be one of those impossible maxwell demon problems 17:17 < kanzure> .wik maxwell's demon 17:17 < CaptHindsight> if it's diluted you just watch for a single molecule to come through and then stop the flow 17:17 < yoleaux> "In the philosophy of thermal and statistical physics, Maxwell's demon is a thought experiment created by the physicist James Clerk Maxwell in which he suggested how the Second Law of Thermodynamics could hypothetically be violated. In the thought experiment, a demon controls a small door between two chambers of gas." — https://en.wikipedia.org/wiki/Maxwell%27s_demon 17:17 < nmz787_i> nah, those ion pumps do it all the time 17:17 < CaptHindsight> you can two filters act as a gate 17:17 < nmz787_i> even polymerase is a sort of nucleotide pump 17:18 < CaptHindsight> or build a polymer that loops 17:18 < CaptHindsight> aluminum oxide is easy to grow and have isolated pores 17:20 < kanzure> hmm so nevermind about it being impossible to isolate single (small) molecules in that way, perhaps it was single leptons or smaller particles 17:21 < nmz787_i> heh, answer... the demon generates entropy, so all is well 17:21 < nmz787_i> I wonder how many scientific articles mention words like demon... or more importantly, wizard 17:21 < kanzure> .gc site:nature.com wizard 17:21 < yoleaux> kanzure: Sorry, that command (.gc) crashed. 17:22 < CaptHindsight> or make a polymer protein hybrid that can act as a filter and gate 17:22 < kanzure> .gc site:sciencedirect.com wizard 17:22 < yoleaux> kanzure: Sorry, that command (.gc) crashed. 17:22 < nmz787_i> 7690 17:22 < CaptHindsight> electrically controlled 17:22 < kanzure> CaptHindsight: there are many membrane-bound proteins that act as filter and gate 17:22 < nmz787_i> CaptHindsight: yep, ion channel 17:22 < nmz787_i> .wik gated ion channel 17:22 < yoleaux> nmz787_i: Sorry, that command (.wik) crashed. 17:22 < nmz787_i> pfft 17:22 < CaptHindsight> so whats the problem then? 17:22 < kanzure> dpk: fixplz? 17:22 < kanzure> CaptHindsight: nobody has demonstrated it 17:22 * dpk kill kill kill Wikipedia 17:23 < nmz787_i> placing the ion channel in the right place/orientation 17:23 < kanzure> dpk: also gc is crashing 17:23 < CaptHindsight> oh you mean in nature there are ion channels 17:23 < CaptHindsight> sure 17:23 < kanzure> indeed, there are many many types of membrane-bound channels 17:23 < andytoshi> kanzure: i minored in physics 17:23 < andytoshi> what's up? 17:23 < nmz787_i> nature pretty much has everything 17:23 < kanzure> andytoshi: single molecule pump. impossible? 17:23 < andytoshi> lol, hmm 17:23 < kanzure> small molecules, not giant dna molecules 17:23 < nmz787_i> proton pumps are like first-year topics 17:24 < andytoshi> hehe nmz787_i 17:24 < CaptHindsight> I'm saying we attach polymers to it to control them 17:24 < nmz787_i> .title http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2742554/ 17:24 < yoleaux> Ion channels versus ion pumps: the principal difference, in principle 17:24 < andytoshi> i am definitely not more qualified than anyone else .. 17:24 < kanzure> qualifications are overrated anyway 17:24 -!- CyberEater [~Genesteal@c-69-255-209-151.hsd1.md.comcast.net] has joined ##hplusroadmap 17:25 < nmz787_i> https://en.wikipedia.org/wiki/Proton_pump 17:25 < kanzure> .wik proton pump 17:25 < yoleaux> "A proton pump is an integral membrane protein that is capable of moving protons across a biological membrane. Mechanisms are based on conformational changes of the protein structure or on the Q cycle." — https://en.wikipedia.org/wiki/Proton_pump 17:25 < nmz787_i> smallest molecule pump 17:25 -!- Darius [~quassel@75.76.19.18] has quit [Ping timeout: 255 seconds] 17:25 < CaptHindsight> graft polymerization onto an ion pump 17:25 < kanzure> protons are not molecules 17:25 < nmz787_i> and all the studies on RNA tugging kanzure 17:25 < nmz787_i> what! 17:25 < nmz787_i> well, I guess atoms aren't molecules 17:26 < nmz787_i> but molecules are atoms 17:26 < kanzure> molecules are atoms? are we sure about this :-) 17:26 < kanzure> they are made of atoms. 17:26 < nmz787_i> anyway, DNA tugging is a great google scholar search, I'm sure ;p 17:26 < kanzure> i'm not sure why i still can't find a single molecule pump in a machine 17:26 < CaptHindsight> quantum movement of base molecules 17:27 < CaptHindsight> kanzure: make one, somebody should 17:27 < nmz787_i> what's that cholesterol pump 17:28 < nmz787_i> brocolli induces it 17:28 < nmz787_i> brocolli 17:28 < kanzure> nanometer-scale molecular pump http://rafiki.tau.ac.il/~rabani/papers/paper44.pdf 17:28 < nmz787_i> broccoli 17:29 < nmz787_i> .wik spp1 17:29 < yoleaux> "SPP1 or SPP-1 may refer to:" — https://en.wikipedia.org/wiki/Spp1 17:29 < CaptHindsight> broccoli induced quantum teleporation of fats 17:29 < CaptHindsight> my next research project :) 17:29 < nmz787_i> jrayhawk: you would probably remember that one... it was a bitter molecule that induced the pump 17:29 < kanzure> what's the gas transition temperature for nucleotides 17:29 < kanzure> or er.. what's the right question 17:30 < nmz787_i> jrayhawk: which I think might have been related to the whole removing old cholesterol before it could oxidize in-vivo 17:30 < jrayhawk> not offhand 17:31 < nmz787_i> ugh, damn gene acronyms 17:31 < CaptHindsight> so nobody has made an electrostatic base gun yet? 17:33 < kanzure> nmz787_i: one way to do a nucleotide pump would be the following.... get a protein that attaches to a single nucleotide, make the protein super massive and large, then manipulate that protein instead of the nucleotide. this can also work for other types of tags. 17:34 < kanzure> also possibly do laser conformational change to release the nucleotide 17:34 < CaptHindsight> a bead for single nucleotide 17:35 < kanzure> oh right, you wouldn't be able to guarantee the reaction 17:35 < kanzure> even after dissosciation 17:35 < kanzure> dissociation 17:35 < CaptHindsight> oh, small pore filters https://www.bu.edu/meller/PDFs/AdvMat18_3149.pdf 17:36 < nmz787_i> kanzure: ion concentration could also work to release the molecule 17:36 < kanzure> CaptHindsight: yes that is the nanopore dna sequencing method 17:36 < CaptHindsight> kanzure: can also isolate single molecules 17:37 < nmz787_i> CaptHindsight: i've thought of using nano wires for programming hybridization areas...until I realized each wire would have to be the size of an atom (so they could hybridize) 17:37 < nmz787_i> possibly using a vacuum and electron gun could work 17:37 < nmz787_i> but then you'd need a reliable way to get the electron to stay in place and not leak off 17:38 < nmz787_i> and also a way to way the pattern with nucleotides 17:38 < nmz787_i> then you'd need to, umm, cryo freeze and add ligase 17:38 < nmz787_i> which also doesn't work in frozen non-aqueous conditions 17:38 < kanzure> "Mycobacterium smegmatis porin A (MspA) is the second biological nanopore currently being investigated for DNA sequencing. The MspA pore has been identified as a potential improvement over αHL due to a more favorable structure.[9] The pore is described as a goblet with a thick rim and a diameter of 1.2 nm at the bottom of the pore.[10] A natural MspA, while favorable for DNA sequencing because of shape and diameter, has a negative core ... 17:38 < nmz787_i> CaptHindsight: the big problem is positioning the nanopore. 17:39 < kanzure> ... that prohibited single stranded DNA(ssDNA) translocation. The natural nanopore was modified to improve translocation by replacing three negatively charged aspartic acids with neutral asparagines.[11]" 17:40 < CaptHindsight> nmz787_i: which problem are you trying to solve? Maybe I'm thinking of something else 17:40 < kanzure> "Another method is the use of nanoelectrodes on either side of a pore.[17][18] The electrodes are specifically created to enable a solid state nanopore's formation between the two electrodes. This technology could be used to not only sense the bases but to help control base translocation speed and orientation." 17:40 < nmz787_i> http://www.ncbi.nlm.nih.gov/pmc/articles/PMC413341/pdf/emboj00076-0052.pdf 17:41 < nmz787_i> The portal protein of bacteriophage SPP1: a DNA pump with 13-fold symmetry 17:41 < kanzure> nmz787_i: here you go, try this http://www.nist.gov/pml/div684/single_092209.cfm 17:42 < kanzure> "The water’s abrupt pressure drop—accompanied by a dash of detergent—breaks its surface tension, splitting it into small droplets. ..... In the microfluidic channel, the water is laced with desired molecules of just the right concentration, so that resulting droplets each pick up on average just one molecule of interest." 17:42 < kanzure> oh... "Inside each droplet, the individual molecules of interest slosh around freely in the relatively roomy sphere" 17:42 < kanzure> so is it just one or not? >:( 17:42 < kanzure> the paper is http://proceedings.spiedigitallibrary.org/proceeding.aspx?articleid=784816 17:44 < kanzure> "Generation and mixing of subfemtoliter aqueous droplets on demand" 17:44 < kanzure> "Surface acoustic waves for on-demand production of picoliter droplets and particle encapsulation" 17:44 < kanzure> http://www.researchgate.net/profile/David_Collins15/publication/240306158_Surface_acoustic_waves_for_on-demand_production_of_picoliter_droplets_and_particle_encapsulation/links/0a85e539a8257e3a77000000.pdf 17:45 < kanzure> probably a piezo could do this too 17:45 < nmz787_i> well it's still just concentration based though 17:45 < nmz787_i> the droplet is not required 17:45 < kanzure> another description http://spie.org/x39621.xml?highlight=x2400 17:45 < nmz787_i> I can't remember if there are materials that only let water pass 17:45 < nmz787_i> as a way of concentrating the droplets 17:46 < nmz787_i> is tyvek water or oxygen? 17:46 < nmz787_i> .wik tyvek 17:46 < yoleaux> "Tyvek /taɪˈvɛk/ is a brand of flashspun high-density polyethylene fibers, a synthetic material; the name is a registered trademark of DuPont. It is often seen used as housewrap, a synthetic material used to protect buildings during construction." — https://en.wikipedia.org/wiki/Tyvek 17:46 < kanzure> "Using our devices, we have been able to produce single droplets with volumes under 1 femtoliter, suitable for performing single-molecule studies" 17:47 < kanzure> .title http://pubs.acs.org/doi/abs/10.1021/ac9014319 17:47 < yoleaux> An Error Occurred Setting Your User Cookie 17:47 < kanzure> "Generation and Mixing of Subfemtoliter Aqueous Droplets On Demand" 17:47 < kanzure> "by means of piezoelectric injection" 17:47 < nmz787_i> hah, they didn't use a global-shutter camera for that paper 17:47 < nmz787_i> the images have motion blur 17:48 < nmz787_i> err, interlacing artifacts 17:48 < kanzure> CaptHindsight: have you ever inkjet printed an emulsion? 17:49 < nmz787_i> kanzure: a flying droplet would dry! 17:49 < nmz787_i> (i'm saying that in a good way) 17:50 < kanzure> wouldn't dry in the right environment 17:51 < kanzure> anyway; i feel a little bit better now that at least one group has done a single molecule isolation system. 17:52 < CaptHindsight> kanzure: sure, that's how we get UV cured inks with thermal inkjet 17:52 < kanzure> ah good 17:52 < CaptHindsight> even easier with piezo 17:53 < kanzure> if your nozzle prints out too many emulsion droplets, you can maybe fix it by moving the head faster (so that they land in different spots anyway) 17:54 < CaptHindsight> aerosol jet 17:55 < CaptHindsight> can you use a low vapor pressure vehicle/solvent? 17:56 < CaptHindsight> then the droplet would not dry as quickly 17:56 < kanzure> CaptHindsight: how do you feel about money being used as a carrot for certain performance/testing targets? :P 17:57 < CaptHindsight> what target? 17:57 < CaptHindsight> and carrot for whom? 17:57 < CaptHindsight> whos money :) 17:57 < kanzure> mine 17:58 < kanzure> well there's a blank section in the doc, so basically i'm figuring out money stuff- there's lots of options, like pay-at-completion, milestones, performance/testing targets, schedule, etc. 17:59 < CaptHindsight> oh for the inkjet? 17:59 < kanzure> yeah 17:59 < CaptHindsight> oh it's mostly for materials 17:59 < CaptHindsight> so it's up front 17:59 < CaptHindsight> for whatever materials 18:00 < CaptHindsight> cut it up in chinks if you want 18:00 < CaptHindsight> we work when there's funding 18:00 < CaptHindsight> whatever you are comfortable with 18:01 < kanzure> yeah i was thinking about that, and i'd be okay with upfront only as long as you grant me the option of having you build something else on the same granty if we can't figure out the chemistry 18:02 < kanzure> or, alternatively, if we can't order the chemistry for some reason (who knows- these people are fickle) 18:02 < nmz787_i> .wik nrec 18:02 < kanzure> s/fickle/absurd and they don't ship to people who are willing to pay 18:02 < yoleaux> "The National Robotics Engineering Center (NREC) is an operating unit within the Robotics Institute (RI) of Carnegie Mellon University." — https://en.wikipedia.org/wiki/National_Robotics_Engineering_Center 18:02 < nmz787_i> pfft 18:02 < nmz787_i> not what I meant 18:02 < CaptHindsight> kanzure: are you concerned about getting the chems? 18:02 < kanzure> 20% concerned? 18:02 < CaptHindsight> what are their hoops to jump through? 18:03 < nmz787_i> for a place like Sigma, you need to show a lease on a commercial property 18:03 < kanzure> nmz787_i: i'm thinking azco biotech 18:03 < kanzure> do we want liquid or solid reagents anyway? 18:04 < CaptHindsight> yeah, I jumped through their hoops years ago 18:04 < kanzure> whoops we can't buy liquid phosphoramidites from azco biotech 18:04 -!- ant4t [~s3an@50.141.79.168] has joined ##hplusroadmap 18:04 < nmz787_i> the POSAM paper said basically 'mix at your own risk (of getting the mix contaminated with O2 or water) 18:04 < CaptHindsight> kanzure: what else do you want to mount on the gantry? 18:04 < kanzure> https://azcobiotech.com/reagents-for-oligonucleotide-synthesis/phosphoramidites/dna-phosphoramidites/ 18:05 < CaptHindsight> I have an account with Sigma 18:05 < kanzure> oh interesting 18:05 < nmz787_i> .title http://probesoftware.com/smf/index.php 18:05 < yoleaux> Probe Software Users Forum - Index 18:05 < kanzure> a sigma account is very helpful 18:05 < nmz787_i> (electron microprobe) 18:05 < CaptHindsight> don't use it since they charge 10-100x what I usually get things for 18:06 < kanzure> i think you might have to spend some time on the phone with these guys to figure out the bottle geometry and cap system, https://azcobiotech.com/cepa-bulk-dg-ibu-20-8110.html 18:07 < CaptHindsight> yeah saw those 18:07 < kanzure> huh, azco biotech claims to offer machine design services too. we should take advantage of that eventually. 18:08 < CaptHindsight> they probably have a 3rd party 18:08 < kanzure> "In 2014 the name of Azco Biotech was purchased by Global DNA Solutions, Inc. a California Corporation. Today Global DNA Solutions, Inc. is doing business as AZCO Biotech" 18:09 < kanzure> nmz787_i: other fun things to use the gantry for if the phosphoramidite chemistr fails us? 18:10 < nmz787_i> lithography 18:10 < nmz787_i> put a bluray laser on the gantry and start whipping out microfluidics 18:11 < nmz787_i> (that version of the machine can come live near me) 18:11 < CaptHindsight> AZCO let me get to the checkout 18:11 < kanzure> so bluray laser microfluidics stuff, or maskless photolithography stuff? 18:11 < CaptHindsight> just some billing info a CC to pay 18:12 < nmz787_i> CaptHindsight: lots of sites will let you get there, then follow up with a phone call 18:12 < CaptHindsight> just a UPS verification of address 18:12 < kanzure> nmz787_i: he has a business at his location 18:12 < nmz787_i> yeah so it could be fine 18:12 < nmz787_i> there is no licensing AFAIK 18:13 < nmz787_i> they mostly just wanted to know that the biz was my biz 18:13 < nmz787_i> or i was part of the company, and that i was allowed to receive chem orders there 18:13 < CaptHindsight> 2 photon polymerization 18:14 < CaptHindsight> lithography and microfluidic fabrication 18:14 < CaptHindsight> I make all the photopolymers as well 18:15 < CaptHindsight> DNA anti counterfeiting marking 18:15 < CaptHindsight> I thought that the POSaM was proven to work years ago 18:16 < CaptHindsight> is there some secret sauce that they didn't mention or are they "pretending" that it works? 18:17 -!- proofoflogic [uid65184@gateway/web/irccloud.com/x-wqcoozsjjgnawftq] has quit [Quit: Connection closed for inactivity] 18:17 < kanzure> it works but there's lots of tweaking of everything 18:18 < nmz787_i> it works but sucks for fidelity and isn't easier than the previous tech 18:18 < CaptHindsight> isn't there a QC method using a CCD for testing each layer? 18:19 < nmz787_i> unless you're interested in arrays for microarray hybridization experiments 18:19 < nmz787_i> where lack of fidelity is usually more tolerable 18:19 < CaptHindsight> doesn't it fluoresce when the oxidation is occurring? 18:19 < nmz787_i> (because of signal to noise I guess) 18:19 < nmz787_i> the deblocking step is orange I think 18:19 < nmz787_i> the trityl group 18:20 < nmz787_i> but that isn't sensitive enough to show if one molecule skipped deblocking 18:20 < nmz787_i> and also the acid can eat away the side of the base, destroying the information-carrying capacity 18:21 < kanzure> there are also other problems like argon or nitrogen blowing away your samples, because they weren't linked to the surface well enough 18:21 < CaptHindsight> kanzure: if you're concerned about what to do with a flatbed inkjet take a look at SGIA :) 18:21 < kanzure> or smearing with fingerprints causing nuclease contamination 18:21 < kanzure> SGIA? 18:21 < CaptHindsight> https://www.sgia.org/ 18:22 < CaptHindsight> t-shirts, posters, banners etc 18:22 < CaptHindsight> printed electronics 18:22 < kanzure> hah, no i was thinking more along the lines of "what about printing yeast and ecoli cells" 18:22 < CaptHindsight> sell it to someone for that application 18:22 < kanzure> oh would they buy? 18:22 < nmz787_i> jrayhawk: I think it was Chris Masterjohn... but I cannot find it using 'chris masterjohn bitter induction' or 'bitter vegetables cholesterol "pump" masterjohn brocolli' 18:23 < CaptHindsight> the dimatix flatbed that size is like $40k 18:23 < kanzure> sigh... i should have guessed. 18:24 < nmz787_i> jrayhawk: I want to think it is close to irk2 or some acronym like that 18:24 < kanzure> oh right, so before i add in the upfront section, i need to figure out the future home for the bugger, i'll try to figure that out today i guess 18:24 < kanzure> nmz787_i: want this thing? 18:24 < kanzure> nmz787_i: would you operate it 18:25 -!- erasmus [~esb@unaffiliated/erasmus] has quit [Read error: Connection reset by peer] 18:27 < nmz787_i> kanzure: hrmm, all the chems having short shelf life seems stress-inducing 18:27 < kanzure> it's a lie 18:27 < kanzure> they are going to be opened in a neutral atmosphere 18:27 < kanzure> don't worry about it, the chemicals are like $9/bottle or something, i can pay that :P 18:28 < nmz787_i> didn't we already have a price list, and it was like $1200 per batch? 18:28 < kanzure> that might have been sigma prices 18:28 < kanzure> you're thinking of the price list from http://diyhpl.us/~bryan/nucleic/fbi-diybio-dna-v1.pdf 18:28 < kanzure> 3rd to last slide 18:29 < nmz787_i> I thought it was Glen Research 18:29 < kanzure> entirely possible 18:29 < nmz787_i> ah, yeah, that's the list 18:29 < nmz787_i> it's clearly a screenshot of google sheets 18:30 < nmz787_i> also the posam paper seemed to list an alternative two-component solvent to take the place of acetonitrile 18:30 < nmz787_i> bbl 18:30 < kanzure> wait you didn't answer 18:33 -!- nmz787_i [~ntmccork@134.134.139.72] has quit [Read error: Connection reset by peer] 18:33 < ParahSailin> is medical marijuana a real thing, or just a backdoor legalization? 18:34 < ParahSailin> second, if its useful for cancer patient relief, how is it generally recommended? 18:34 < ParahSailin> take it during a chemo session? 18:35 < ParahSailin> or only the rest of the time 18:35 < CaptHindsight> kanzure: http://www.myprintresource.com/product/10208199/fujifilm-north-america-corporation-fujifilm-dimatix-dmp-5000 no longer in production 18:36 < CaptHindsight> only 500 x 500 mm print area and that one was closer to $100K 18:37 -!- CyberEater [~Genesteal@c-69-255-209-151.hsd1.md.comcast.net] has quit [Ping timeout: 256 seconds] 18:37 -!- CyberEater [~Genesteal@c-69-255-209-151.hsd1.md.comcast.net] has joined ##hplusroadmap 18:38 < kanzure> CaptHindsight: don't the $200k machines come with support contracts etc.? 18:38 < kanzure> and on-site maintenance contracts 18:38 < CaptHindsight> they come with whatever they get you to pay for 18:38 < CaptHindsight> that is usually above the price of the printer in the fine print 18:39 < CaptHindsight> or they lease it to you with a service contract built in 18:39 -!- proofoflogic [uid65184@gateway/web/irccloud.com/x-qxwacvqkttuiwzxc] has joined ##hplusroadmap 18:40 < CaptHindsight> and they watch you like a hawk to see if have done anything like use a 3rd party ink 18:40 < CaptHindsight> so you void the service plan 18:40 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has joined ##hplusroadmap 18:43 < kanzure> ah interesting 18:43 < kanzure> it's about the same with dna synthesizers for that matter 18:43 < CaptHindsight> does it still need a home? 18:44 < kanzure> possibly- nathan ran off 18:47 < CaptHindsight> any private rooms at the nearby hackerspace? 18:48 < juri_> i imagine BUGS would enjoy it. 18:49 < juri_> and i can get there via the metro+bus, to operate. 18:51 < kanzure> juul would probably take it at counterculture labs 18:52 < CaptHindsight> https://counterculturelabs.org/ wow 18:52 < kanzure> superkuh: do you want it? 19:04 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has quit [Ping timeout: 264 seconds] 19:08 * superkuh reads the backlog. 19:10 < superkuh> No. 19:14 -!- knobuddy [~Razk@unaffiliated/knobuddy] has joined ##hplusroadmap 19:24 -!- erasmus [~esb@unaffiliated/erasmus] has joined ##hplusroadmap 19:27 -!- ant4t [~s3an@50.141.79.168] has quit [Ping timeout: 240 seconds] 19:33 -!- Viper168 [~Viper@unaffiliated/viper168] has quit [Quit: Leaving] 19:35 -!- Viper168 [~Viper@unaffiliated/viper168] has joined ##hplusroadmap 19:44 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has joined ##hplusroadmap 19:54 < nmz787> bak 19:57 -!- CyberEater [~Genesteal@c-69-255-209-151.hsd1.md.comcast.net] has quit [Quit: Leaving] 19:57 -!- genehacker [~chatzilla@8-44.ptpg.oregonstate.edu] has quit [Ping timeout: 246 seconds] 19:57 -!- Manti5 [~Genesteal@c-69-255-209-151.hsd1.md.comcast.net] has joined ##hplusroadmap 19:58 -!- Manti5 [~Genesteal@c-69-255-209-151.hsd1.md.comcast.net] has left ##hplusroadmap [] 20:07 < kanzure> nmz787: you haven't answered either 20:18 -!- PatrickRobotham [uid18270@gateway/web/irccloud.com/x-knzxqmqtwcqwsjcp] has joined ##hplusroadmap 20:20 < nmz787> kanzure CaptHindsight: why not aim for a sub-$500 CNC with 0.1-0.5 uM positioning repeatability, using interferometry (requiring, most likely, to either utilize the optics in the drives somehow (it /might/ be possible) or injection molding for plastic optics) or machine vision using a grid-recticle (but I imagine the feedback would be quite slow) 20:20 < kanzure> for the dnajet? 20:21 < nmz787> is that price-point not possible for that accuracy? in production, or rather once production can be achieved (through prototyping, making molds, etc) 20:21 < nmz787> it seems like if you could make a smaller one with more accuracy, if making them smaller would make the machine cheaper 20:22 < nmz787> then you could have a usable thing for litho, pipetbot, etc... and then go on to prototype dnajet chem 20:22 < nmz787> or straight to it 20:22 < nmz787> i don't think more than 25cm x 25cm machine is necessary (that's an 8 inch wafer) 20:23 < nmz787> (actually 8 inch is 20.32 cm) 20:24 < nmz787> the mesa cards seem quite expensive 20:24 < kanzure> if you don't want this thing, just say so 20:24 < nmz787> but that can be reduced later, and keep the fabric right? 20:24 < nmz787> or HDL? 20:24 < nmz787> verilog? 20:24 < kanzure> i can ship it to someone else 20:24 < kanzure> it's vhdl stuff 20:24 < kanzure> actually it's probably verilog but i hope it's systemc. 20:25 < nmz787> would this fit on an ice40 CaptHindsight or juri_ ? if not, how undersized is an ice40? 20:25 -!- Darius [~quassel@75.76.19.18] has joined ##hplusroadmap 20:25 < kanzure> (sleepmode) 20:26 < nmz787> kanzure: I don't think I have time to do the chem for the machine right now 20:26 < CaptHindsight> then you have to make boards, cables etc 20:26 < kanzure> nmz787: you wouldn't have to do the chemistry 20:26 < nmz787> kanzure: it is hard for me to make time, as the value proposition doesn't seem like a good ratio of effort to payoff 20:26 < kanzure> nmz787: although you would be asked to operate it (turn it on) and perform maintenance (like switching out chemicals) 20:26 < nmz787> or rather, the best ratio 20:26 < kanzure> and wifi config stuff 20:27 < nmz787> who will do the chem debug then? 20:27 < kanzure> 1.2 million spots/chip is not good ratio of effort to payoff? 20:27 < CaptHindsight> drivers, maybe a new CNC motion application.... 20:27 < nmz787> yeah but the spots are not that great afterward... you still have tons of effort left 20:27 < kanzure> nmz787: i think captain is going to be doing the chemistry debugging 20:28 < nmz787> if it's "turn the thing on, and load chemicals" then it could sit here... but I think one of the biohacker spaces would have far more effort freely available that would be interested 20:29 < nmz787> I imagine 20:29 < nmz787> have you asked keoni? 20:29 < kanzure> keowho? 20:29 < nmz787> koeng 20:29 < kanzure> keoni gandall? 20:30 < kanzure> don'tk now this person 20:30 < nmz787> yeah 20:30 < nmz787> he has a bio-internship for a while now at UCLA I think, and hangs out with L.A. biohackers 20:30 < kanzure> oh the los angeles people, yeah i guess we could ask them 20:30 < kanzure> cory would be a good home for this 20:31 < nmz787> or maybe it's irvine 20:31 < nmz787> yeah 20:31 < nmz787> UCI 20:32 < kanzure> ok just sent an email to cory 20:33 < kanzure> jrayhawk: you want to run this? 20:35 < kanzure> fenn: also haven't asked you yet 20:36 -!- Madplatypus [uid19957@gateway/web/irccloud.com/x-ktdebwyjroxkwhns] has quit [Quit: Connection closed for inactivity] 20:39 < kanzure>  20:39 < kanzure> jojack might be a good person to keep this, although i wouldn't expect him to do chemistry 20:44 < jrayhawk> Run what with the who now? 20:55 -!- Viper168_ [~Viper@unaffiliated/viper168] has joined ##hplusroadmap 20:57 -!- Viper168 [~Viper@unaffiliated/viper168] has quit [Ping timeout: 252 seconds] 21:12 -!- Quashie [~boingredd@50.14.92.17] has joined ##hplusroadmap 21:18 -!- Quashie [~boingredd@50.14.92.17] has quit [Ping timeout: 264 seconds] 21:19 -!- knobuddy is now known as JimmyRussel 21:20 -!- JimmyRussel is now known as JamesRussel 21:20 -!- JamesRussel is now known as JimmySRussel 21:24 -!- Madplatypus [uid19957@gateway/web/irccloud.com/x-dvyjjsdompilywnq] has joined ##hplusroadmap 21:38 -!- Viper168_ is now known as Viper168 21:44 -!- Darius [~quassel@75.76.19.18] has quit [Remote host closed the connection] 21:49 -!- nottimschmidt [~timschmid@2601:405:4101:52db::822] has quit [Ping timeout: 248 seconds] 21:53 < kanzure> jrayhawk: dna synthesis equipment 22:04 < jrayhawk> i do not have a fume hood, no 22:05 < jrayhawk> if you want to buy me a fume hood, maybe 22:06 < jrayhawk> i have like zero lab experience; i suspect diybio folks would be a saner choice 22:25 < nsh> .t http://www.wired.com/2015/07/paradoxical-crystal-baffles-physicists/ 22:25 < yoleaux> nsh: Sorry, I don't know a timezone by that name. 22:26 < nsh> .title 22:26 < yoleaux> Paradoxical Crystal Baffles Physicists | WIRED 22:26 < nsh> -- 22:26 < nsh> The material, a much-studied compound called samarium hexaboride or SmB6, is an insulator at very low temperatures, meaning it resists the flow of electricity. Its resistance implies that electrons (the building blocks of electric currents) cannot move through the crystal more than an atom’s width in any direction. And yet, Sebastian and her collaborators observed electrons traversing orbits millions of atoms in diameter inside the crystal in response to 22:26 < nsh> a magnetic field—a mobility that is only expected in materials that conduct electricity. Calling to mind the famous wave-particle duality of quantum mechanics, the new evidence suggests SmB6 might be neither a textbook metal nor an insulator, Sebastian said, but “something more complicated that we don’t know how to imagine.” 22:26 < nsh> -- 23:06 < fenn> yep PCR barcode is meant to select a subset of the synthesis pool so it should have unique and maximally distant sequences 23:07 < fenn> i am personally not interested in running this dna synthesis machine and i don't have the facilities for it either 23:09 < fenn> i dont get why you're still talking about single molecule pumps 23:15 < fenn> eventually you will reinvent the ribosome 23:21 -!- Acty [uid89656@gateway/web/irccloud.com/x-sozxtwlywxurstxe] has quit [Quit: Connection closed for inactivity] 23:29 < fenn> if dna synthesis doesnt work out, the inkjet system could be useful for printing electronics; organic semiconductors and 3d circuit boards 23:37 -!- proofoflogic [uid65184@gateway/web/irccloud.com/x-qxwacvqkttuiwzxc] has quit [Quit: Connection closed for inactivity] 23:38 < fenn> fwiw counterculturelabs is right down the road from me and i sort of know kathy buehmann already (and juul i guess from this channel) 23:50 < fenn> if it's nearby i would be willing to get the machine set up in its new home and do some process debugging --- Log closed Tue Jul 14 00:00:10 2015