PCR Reamplification

ww40 William_D_WARREN at UMAIL.UMD.EDU
Wed Jun 3 13:56:00 EST 1992


>Having heard some of the many suggested ways to reamplify a pcr 
>product I thought I would let you know how we do it.
>First i am assuming you are trying to amplify using the same primers.
>1 Make a mix in a pcr tube without any template.
>2 Take pcr tube with product that needs to be reamplified.
>3 Take 1-2ul (any small amount will do) and put in the the new
>tube as if it were the template.(which it is :-))
>4 Put tube in pcr machine and that's it.
>Note NO purification at all. (total waste of time)
                            ^^^^^^^^^^^^^^^^^^^^^^^^^
I disagree strongly here ... there are some applications where
particular combinations of primers, degenerate primers, and/or low
annealing temperatures are used and PCR band purification is quite
necessary (if not manditory) prior to a reamplification.

Bill Warren
Center for Ag Biotech
University of Maryland




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