Library amplification problems - Glenn Samwell

Tue Feb 4 19:29:53 EST 1997

Hi yeast users,

I am trying to amplify a human brain library (Clontech) that has been 
cloned into pACT2 and transformed into E. coli strain BNN132. The yields 
I have been getting are very poor and I think that it is because the 
bacterial cells are not lysing properly. I am using standard maxi-prep 
protocols and I'm not sure what else to try. I thought maybe a couple of 
freeze-thaw cycles may assist in cell lysis. Has any body had the same 
problems, and what, if any, methods did you use to get better yields?

Thanking you in advance,


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