I've said this before, but, basically, i've been using a PCR reaction with Pfu on genomic DNA, and until very recently, it's worked really well. last few weeks, the reaction flopped majorly. Other researchers in our lab have used the Pfu also, unsuccessfully. So, we got a new batch out of our freezer, it too didn't work. Changed the water... didn't work. Fresh dNTP... didn't work, fresh primers... didn't work. changed the PCR machine from my floor to the one upstairs... didn't work. swore... didn't work. changed from pfu to another proofreader with it's very own buffer and with assurances that this PCR would work... didn't work (though did get some lovely primer dimers)....
I was wondering... and was hoping someone could confirm this.... the only thing that has remained constant is my supply of DMSO (5%). could this be the thorn that screwing me around? (as i write this, i'm running a PCr with a fresh batch of DMSO)
can DMSO go bad?
hoping desperately that this reaction works.
supposing this reaction doesn't work, and i manage not to throw myself off a building, is there anything else i can try. this reaction has worked for months, and now is not.
fresh DMSO... didn't work. i don't get it. I got this reaction to work, One day it's fine... next it's stuffed. Which molecular biology god should i make a sacrifice to?
Edited by vetticus3, 01 December 2004 - 07:53 PM.