PEG transformation of E. coli???

bchs1b at jetson.uh.edu bchs1b at jetson.uh.edu
Tue Oct 2 22:14:05 EST 1990


There is a procedure for the transformation of E. coli using PEG in the
Current Protocols manual (I forgot the original reference) which supposedly
gives nearly the same efficiency as CaCl2 but is somewhat simpler. We have
tried this a few times in my lab and have gotten 0 tranformants while with
the same cells and DNA using CaCl2 gotten our normal pretty high level of
transformation. I am pretty baffled as to why a simple procedure absolutely
failed to work in our hands. We tried making the solution up a few times
using at least 2 different batches of chemicals. 

Has anyone successfully used this procedure? Was some important step left
out? Are we being bozo's somehow? Although our lab has not crashed because
of this, I certainly would like to know what is going on. Any comments or
suggestions?

Mike Benedik
biochemical sciences
university of houston                           benedik at uhou (bitnet)
houston, TX 77204-5500				benedik at uh.edu



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