Excerpt from protocol/labnotes

Excerpt from the protocol:

Nenufar – antiviral research project 01 – Expression and purification of peptide LBOUNCER from the synthetic  gene VRBNCR-01 inserted into an e-coli plasmid vector, transformed and induced to grow on bacteria.  Explore how to do this in a fast and consistent way with the highest purity.

Experiment 01A:

DNA synthesis of the VRBNCR-01 gene which is inserted into a pET15 expression plasmid with a His 6 tag on the N-Terminus followed by a thrombin cleavage site.  The gene will be cloned into pET15b via NdeI and BamHI. We will use heat transform into competent cells, then plate out on Amp-agar to grow colonies.  Induce for expression.

Then on to Experiment 01B: purification using chromatography and SDS page

I contacted a lab in the San Francisco Bay Area where we can work.  All in all, we are going to do a lot of improvising so we can stretch the budget and work for at least 6 months on the experiment. Everyone is volunteering their time. We have wonderful mentors and we are learning by doing, the biohacker way.  It is always a steep learning curve as anyone who has worked in molecular biology knows.  The journey of discovery makes it worthwhile.  Thanks to all of you who are making this possible.