PCR primer storage?
biomedlover at hotmail.com
Thu Mar 28 12:25:10 EST 2002
In article <c22e31f9.0203260346.5605154c at posting.google.com>,
bonehammer73 at yahoo.com says...
> acoustic52 at hotmail.com (NM) wrote in message news:<410c9a77.0203231709.1f72b270 at posting.google.com>...
> > What is the best diluent to use to dilute PCR primers? ANy
> > advantage/disadvantage to storing them at -70??
> To simplify our lives, in my lab we prefer not to add any salt to the
> PCR other than those already present in the PCR 10x buffer. Thus, to
> dilute primers, we use distillated, autoclaved water and make small
> 100 uM aliquots ("mothers").
> From these, we prepare 25 uM working solutions, also using distillated
> water. Mothers can be kept at -70 with no problem whatsoever, working
> solutions go together with the rest of PCR reagents at -20 (*far away*
> from the DNA)
> Chiara Martellossi
In our lab, the stock primer is 400uM, and work primer is 10uM. For every
25 volumn PCR reaction, we use 0.3ul per single PCR. BTW, we use the
distillated, autocaved water to resolve the primer powder after being
gotten from the company. The stock one is stored in -80 degree frig, and
work one in -20 degree frig.
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