D.J.Studholme at soton.ac.uk
Thu Nov 30 05:47:37 EST 2000
In response to Rene's question about transforming B. stearothermophilus - I had big problems
transforming this bug a few years ago (and in fact appealed for help in this newsgroup and recieved some very helpful replies).
Anyway, I found the method that worked best was Issay Narumi's electroporation method (published in
Biotechnology Letters, around 1993, I think). There is great variation in efficiency of electrotransformation between different
strains of B. stearothermophilus - it worked very well with Dr. Narumi's strain K1041. In fact it turns out that this strain
is actually Bacillus thermodenitrificans rather than B. stearothermophilus - but this may or may not not be a problem for you.
As a positive control for the transformation DNA you can use pUB110 DNA which you can buy from Sigma-Aldrich. This confers
Kanamycin resistance. Also the methylation state of the DNA seems to be important- so be careful if you use any shuttle vectors that have been passged through E. coli.
David J. Studholme, PhD.
Division of Biochemistry & Molecular Biology
School of Biological Sciences
University of Southampton
Bassett Crescent East
djs at soton.ac.uk
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