From Counter Culture Labs
DIY Taq polymerase
- One is to make your own Taq enzyme for PCR. I have the construct and the protocol but would need a few special items to do the isolation. Anyone interested?
- i might be able to help you with that Ramon. though usually bacterial transformation is done in a lab - does are there any legal issues with doing it somewhere besides a lab ? I just don't know so I'm asking.
- It has been a long time since I have done this but my recollection is that the plasmid construct I have is very different from the open source one. Also, the Taq I am trying to make comes with a His tag so we can purify buckets of the stuff. I already have glycerol stocks of bacteria and would need IPTG and maybe some columns to purify.
- Correction, No HIS-tag on the Taq so we need to get a columne with Bio-Rex 70 resin, bead size 150-300, mesh size 50-100.
- The open source vector is inexpensive and no his tag (as you mentioned) so would not need to purchase potentially expensive nickel columns - only need the IPTG. Any interest in purchasing the open source vector? i have a protocol for purifying taq using that vector if needed - lyse cells and heat. Actually the protocol is on openbiotech.