Enhancement of PCR?

Richard Hastings mbxrh at unicorn.ccc.nottingham.ac.uk
Mon Mar 6 08:04:18 EST 1995


I've recently been having some problems with contamination of PCR, i've
had some advice as to how to minimise contamination and smearing.  One
such piece of information was to boil the PCR prmers for 5 minutes then
snap freeze in liquid nitrogen and thaw on ice before using in the PCR
reaction.  This apparently reduces smearing of products, i found that
this treatment also enhanced the PCR reaction somewhat, producing
stronger bands in both my main reaction as well (unfortunately) as my
no template negative control.
I generally store aliquoted stocks of primers at -20C then take an
aliquot and store at 4C and use that rather than refreezing each time
(which is supposed to have a detrimental effect on the primers).
Does anybody know why this enhancement is occuring?
Thanks for any help.

Richard Hastings (mbxrh at unicorn.nott.ac.uk)
Dept. of Biochemistry,
Queens Medical Centre,
University of Nottingham,
Nottingham NG7 2UH.



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