Project ideas

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DIY Taq polymerase

Ramon Mendoza

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?

Ron Shigeta

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.

Ramon Mendoza

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.

Nina DiPrimio

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.

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