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smears in PCR products (bisfulite treated DNA)

Started by sri2010, May 12 2011 11:15 AM

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#1 sri2010

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Posted 12 May 2011 - 11:15 AM

Hi Everyone,

I've successfully amplified my products with:

Zymo research's "human methylated DNA".

However, when I extract DNA from cell culture, I don't see any products- only smears. I've designed my primers using methylprimer express.

I do 40 cycles.

What does this mean?

Thanks.

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#2 pcrman

Epigenetist

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815 posts

Posted 15 May 2011 - 08:56 AM

You got expected products using control templates, suggesting the primers work. Make sure your DNA quality (purity) and recovery are good. You may also try hot start taq polymerase such as JumpStart redTaq from sigma.

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#3 sri2010

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Posted 17 May 2011 - 07:36 AM

pcrman, on 15 May 2011 - 08:56 AM, said:

You got expected products using control templates, suggesting the primers work. Make sure your DNA quality (purity) and recovery are good. You may also try hot start taq polymerase such as JumpStart redTaq from sigma.

I've tried that but no luck. Only smears...

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