help PCR

Björn Rosén bjornrosen at swipnet.se
Mon Oct 20 11:40:09 EST 1997


Tom wrote:
> 
> jkennie at GPU.SRV.UALBERTA.CA (J Kennie) wrote:
> 
> >On 1 Oct 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

___________

Hi, this is a reproducibility problem. If you need advice contact me at
my email address (bjornrosen at swipnet.se).

Regards, B. Rosen



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