Blunt ended PCR products?

Richard Sucgang rs54 at cunixf.cc.columbia.edu
Fri Mar 13 12:23:52 EST 1992


In article <1992Mar12.172225.293 at bio.aau.dk> brian at bio.aau.dk writes:
>Hi out there!!!
>
>I got a problem (who haven't).
>
>Has anyone expirence in getting blut ended PCR products.
>I've read in a resent issue of PNAS that there was succes when VENT polymerase
>was used. Any comments???
>If I treat the PCR product with klenow, do I then have to heat the samples
>to ~99 degrees for 10 min to inactivate the TAQ-pol??? (this precedure is 
>not good since I have to products with the same sequence, but one of them has
>a deletion, and if the samples are heated I will get mixed products)
>
>I hope someone has a suggestion, if so please mail.
>
>Peter Kristensen
>Department of Chemistry
>division of Biostructural Chemistry
>Aarhus
>Denm

Well, why don't you just EtOH ppt before the Klenow step?
Dat's wot I do...


-rich
Richard Sucgang : Dept. of Anatomy and Cell Biology
Columbia University (sucgang at cuhhca.hhmi.columbia.edu; 
de slime god         rs54 at cunixf.cc.columbia.edu)



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