preparation of glycine competent cells..

B.Rama chandran via methods%40net.bio.net (by chandranbrama from gmail.com)
Sat Jan 30 08:44:29 EST 2010


Respected Sir/Madam,
       I am facing problem in preparing Glycine competent cells ( *E.coli*
BL23 DE3 ) with good efficiency.
       I am using freshly prepared Glycine, transformation buffer, DNA
(quality is 260/280m is 1.89). Cells were harvested OD (600nm)  between
0.9-1.0. But  I am getting only few colonies (10-20 colonies ). For protein
purification I need a plateful colonies. Please give me some suggestion.

Yours sincerely,
B.Ramachandran.

Project Assistant,
MCBL, IISc,India.


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