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Primers too dilute

primers PCR

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#1 stickperson22



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Posted 18 April 2012 - 08:19 AM

I am performing PCR with primers on a gene that is about 10kb long. I am not getting any results from my gel, all I see is a smear. I believe that my primers are too diluted, but I am not sure how to go about solving this. Anyone have any ideas? Thanks!

#2 hobglobin


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Posted 18 April 2012 - 08:26 AM

What concentration you use then? Stock solutions usually have 100 pmol/microl and working solutions e.g. 10 pmol/microl that are then diluted to changing final concentrations in the PCR mix.
And for 10 kb you perhaps to try a long range PCR approqach that many companies offer (if you're not using it already)...

One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that did belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

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