# Key Technical and Conceptual Points from IRC Discussions

• Participants discussed advanced genetic engineering techniques, specifically methods for inserting PCR'd DNA into plasmid vectors using Pichia pastoris yeast as an expression system, with particular focus on using secretion tags to have cells export proteins into culture medium for easier harvesting.

• A detailed microfluidic chip design was explored, featuring channels for storing DNA droplets with calculations suggesting 20,000-25,000 drops would require approximately 50mm of zigzag or spiral channel length, with technical specifications including 20-micron channel width and 2.5-micron wall thickness.

• The group analyzed various distance sensing technologies for precision pumps, comparing optical encoders, laser-based systems, and camera-based volumetric measurement for achieving nanoliter precision in fluid handling.

• A DIY retinal projection system (http://eclecti.cc/hardware/blinded-by-the-light-diy-retinal-projection) was referenced, which projects images directly onto the retina, though limitations were noted regarding wide-angle image creation.

• One participant described work on producing spider silk proteins using Pichia yeast in bioreactors, discussing growth media options and the challenges of protein polymerization, which requires specific pH changes and extrusion pressure to form fibers.

• The conversation highlighted reproducibility issues in biological research, noting that many procedures work inconsistently ("try 20 times, it works once"), underscoring standardization challenges in open-source biohacking.

• References were made to biohacking communities and resources, including Copenhagen's Biologigaragen, Labitat, and Copenhagen Suborbitals, as well as TX-RX hackerspace in Houston.