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PCR product biotinylation


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#1 krildrik

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Posted 19 December 2012 - 06:07 AM

Hi everybody!

Have anybody done PCR product biotinylation? Can anybody share protocols of doing it? I am trying to uderstand this article http://www.sciencedi...890850804001008 , particulary why they use biotinylated Forward primers, if you can do it just by incorporating already biotinylated dUTP while PCR for example. I would be really grateful if you share some thoughts or info.

#2 phage434

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Posted 19 December 2012 - 02:23 PM

Yes, you can incorporate biotin-dUTP by PCR in any reaction. Simply add some biotin-dUTP to the dNTP mix (10% or so of the dTTP concentration or so). Use Taq as the enzyme -- the better modern enzymes will not incorporate dUTP.

#3 krildrik

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Posted 20 December 2012 - 12:08 AM

Thaks a lot phage434 , but maybe you can glimpse this article because this bitinylated Forward primer just blows my mind.

#4 phage434

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Posted 20 December 2012 - 05:34 PM

Sorry, I don't have access to the article.




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