Transformation Opinions!

Jeffery Haines jhaines at uoguelph.ca
Sat Aug 11 14:12:19 EST 2001


Hello all!

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!
Jeff

PS:  Plasmids are being transformed into RbCl comp. cells


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~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o~o
Jeff Haines
Undergrad. in Dept. of Chem. and Biochem.
University of Guelph.




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