preparation of glycine competent cells..

[B.Rama chandran]

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.