fast cloning

D.K. dk at no.email.thankstospam.net
Sun Aug 18 13:03:21 EST 2002


"aes" <aes at donnotreply.com> wrote:
>Hi.
>
>The few years that I have been cloning I have always used standard
>procedures for transformation (i.e 15 min. on ice after addition of
>plasmid/lig.mix followed by heatshock, ice again, more LB and 1 hour of
>recovery). I have just heard of other people that simply mix competent cells
>with plasmid and incubate at RT for 10 min. before plating. Is this another
>standard procedure and is it much more inefficient??
>

This will only work with ampicillin selection. With others, you have to allow
for a time to synthesize appropriate gene product. 

Relative efficiency of rt vs heat shock probably depends on the nature 
of competent cells. Our Inoue cells give more colonies w/o heat shock,
standard Ca cells (I am told) work better with shock. 

DK



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