SAP eats my vector

Sasha Kraev bckraev at aeolus.ethz.ch
Thu Dec 15 05:29:32 EST 1994


James, why on earth are you using this (expensive) enzyme, if you are to gel
purify your vector!? Just use "normal" CIAP, run the reaction on an 1% LGT gel,
cut out the band, digest with gelase and ligate: almost zero background is 
what you get. I do not know, where CIAP goes on agarose, but the gel box 
never gets contaminated with it. I never bought the trick of heat inactivatable
phosphatase, since it is the uncut vector that you have to get rid of anyway 
( by gel purification ). Can post details of the procedure, if required.
Good luck.



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