topo ta cloning
hzhen at freeuk.com
Thu Dec 30 14:44:30 EST 2004
> Hi again.
> Thanks for the answers.
> I decided to test the difference between recovery and no recovery.
> So I transfected 50 µl of invitrogen top10 cells with 1µl of ligation mix,
> and after heatshock I plated 25µl on amp plates and the other 25µl I
> incubated with SOC at 37 for 40 min.
> On the first plate I got around 30 colonies and on the other plate, around
> 300. So I guess it is worth doing after all!
The difference might be smaller than you think. At an optimum growth
rate of around 20 minutes doubling time, only a quarter of the 300
colonies you see may actualy be transformants. Personally I found the
differences to be minor, and normally unimportant unless the ligation
you do is difficult.
> Happy New year.
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