PCR questions...

knud kknald at hotmail.com
Tue Apr 2 03:31:01 EST 2002


1) It is unlikely that your MgCl solution has "gone off", by the way you do
not have to freeze your MgCl solution but remember to give it a good shake
before every time that you use it. If you want to freeze the solution it is
allright, no damage is done.
2) It depends on the polymerase you are using. If you are using a normal taq
polymerase a hot start is not recommendet. If your polymerase is a hotstart
polymerase you'll have to hotstart for 15'  to activate the polymerase.

Mads hollegaard


"NM" <acoustic52 at hotmail.com> skrev i en meddelelse
news:410c9a77.0203312330.50968585 at posting.google.com...
> Two questions:
>
> 1) Will MgCl 'go off' after repeateded freeze/thaws??
> 2) Is it common to do a PCR without doing a hot start and see nothing,
> but then get a great product using a hot start??? It seems unlikely
> that it would go from nothing to something under these conditions...
> by the way, the product size is 1100bp...





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