pcr conditions

Steven L Goldberg steven.goldberg at bms.com
Tue Apr 25 07:21:57 EST 2000


Try one of the kits designed to optimize your reactions (I like FailSafe
from Epicentre or OptiPrimer from Stratagene) and use denature 94oC, anneal
50oC, extend 72oC (1 min/kb) as a starting point.  If your primers are
prepared correctly and hybridize to your template, you will almost always
get what you want.  You can then tweak the annealing temperature and other
conditions to maximize the desired product if needed.

Karen Roberts wrote:

> Hi all,
>
> I don't know a whole lot of pcr so this question is probably a little
> basic.
>
> How are pcr conditions determined?  the pcr rxns I have done in the past
> always was someone giving me the cycling conditions.
>
> Is there a formula of some kind?  I am having trouble on some pcr and
> I'd like to address this potetial source.
>
> TIA,
> Karen





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