nested pcr

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shuk08
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nested pcr

Post by shuk08 » Tue Oct 06, 2009 12:29 pm

How to get rid of larger non-specific bands appearing in 1st round of pcr just above the expected bands (of approx. 1.5 kb)? The non-specific bands from the 1st pcr are amplified in 2nd round of pcr using nested pair of primer.
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JackBean
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Post by JackBean » Tue Oct 06, 2009 2:02 pm

Are you asking or just saying us?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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canalon
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Post by canalon » Tue Oct 06, 2009 2:19 pm

I would try to optimize the first rund of PCR. Tips and tricks to do thta can be found here:
http://info.med.yale.edu/genetics/ward/tavi/Guide.html
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)
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roniadam
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Post by roniadam » Sat Oct 10, 2009 9:15 am

I had the same problem before. I advice you to use an adjuvant to improve your PCR effeciency. I used Q solution that comes with hotstar PCR master mix and it gives me a very good amplicon.
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