I'm not sure I understand what you are doing. Really, for recombinant
protein expression you only would need to recover a single colony
containing your construct. People typically grow liquid cultures to
produce recombinant proteins.
If you are including IPTG in your plates (and it sounds like you
might be) I would suspect this might greatly reduce transformation
efficiency.
Mike
On Jan 30, 2010, at 7:44 AM, B.Rama chandran wrote:
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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