jhaines at uoguelph.ca
Sat Aug 11 14:12:19 EST 2001
I have a question about transformation. The lab I'm working in
recently purchased a new E.coli strain from Stratagene (their new BL21
cells). Anyways, they recommend making up SOB medium, adding sterile
glucose (then it becomes SOC medium, I think), and transforming their
cells with SOC *and* B-mercaptoethanol.... talk about a pain in the neck!
I've tried it once, and it is a long and tedious procedure -- and they
say to make up the SOC fresh every time. I know this is what
Stratagene recommends, but honestly, is this absolutely necessary? (I
would prefer to use the "add plasmid, incubate on ice, heat shock, add
LB, mix 1 hr, plate). So essentially, the question is: Does it make a
difference for cell "health" (or further down the road, protein
expression) depending on what protocol one uses?
What would be the purpose of B-mercaptoethanol anyways?
Thanks so much!
PS: Plasmids are being transformed into RbCl comp. cells
Undergrad. in Dept. of Chem. and Biochem.
University of Guelph.
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