help PCR

Mario Chin user at grgs.com
Wed Oct 1 13:03:32 EST 1997



Julian Robinson wrote:

>       Iam trying to amplify a ~450bp fragment with two 20bp primers.
> It was working perfectly before I went on holiday. However when I
> came back I begtan to have problems. The problem was smears in all
> the lanes, including the control lane. It wasn't the template DNA
> that was the problem (it oocured in the -ve control), and it is'nt
> contamination (new everything has been used).
> We are leaning towards primer dimers as the answer at the moment (as
> there is some complementarity? between the primers), but the real
> question is why has it just started to occur, when it was perfect
> (i.e. no smears and clean bright product) before I went on holiday?
>
> many thanks
> julian
> ---
> julian.robinson at plant-sciences.oxford.ac.uk


If the -ve control also have smear like the other lanes, the template is
probably degraded.
--
de mario

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