PCR from genomic dna

Roland Hübner rhubner at gins.uia.ac.be
Tue Jun 13 14:40:14 EST 2000


Henri:
>>> The amount of genomic DNA we use in a typical PCR is normally about 
>>> 300 ng. We find out that addition of 10 % DMSO to the PCR mixture
Roland:
>> what Mg++ is used with 300ng nucleic ac.? 
Duncan: 
> Will that make that much of a difference? If you were taking free dNTPs
> then I would say it matters.

 Well, when the concentration of dNTPs or primers is altered, the 
concentration of free Mg++ is affected...
 I noticed that it is quite common to find an EXTRA excess of primers in 
such template-"loaded" PCRs, but often the Mg++ is not adjusted...
 
Roland






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