PCR questions...

knud kknald at hotmail.com
Fri Apr 5 16:02:22 EST 2002

I'm sorry if I caused any confusions in my previous answer. The 15'
activation time is only use when you are using the Qiagen Hotstart
polymerase. Always REMEMBER to read the instructions given with the
polymerase to ensure proper handling.
I thank Knud Sorensen from R&D Sigma Aldridge, USA, for making a personal
notice to me about this-

"knud" <kknald at hotmail.com> skrev i en meddelelse
news:3ca96dd3$0$32894$edfadb0f at dspool01.news.tele.dk...
> 1) It is unlikely that your MgCl solution has "gone off", by the way you
> 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
> allright, no damage is done.
> 2) It depends on the polymerase you are using. If you are using a normal
> polymerase a hot start is not recommendet. If your polymerase is a
> 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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