help TA cloning!

EK khatipovNO-SPAM at NO-SPAMuchicago.edu
Thu Dec 19 17:57:00 EST 2002


"Wolfgang Schechinger" <hubahopp at gmx.de> wrote in message
news:3.0.6.32.20021218103436.0081fca0 at pop.gmx.net...

...
> - digest vector with the same 2 favourite enzymes and include some SAP(aso
> in one RxN)
> - spin column purify vector and insert, EtOH prec.
> - ligate, transform, check.
>
> Usually works like a charm, except on mondays past full moon.
>
> Wo
>
Would adding SAP really make any difference? I always thought
dephosphorylation is not necessary if you digest with 2 enzymes that produce
non-compatible overhangs? SAPing would certainly be necessary in case of 2
blunt cutters though.
Emir





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