PCR product biotinylation - (Dec/19/2012 )

Hi everybody!

Have anybody done PCR product biotinylation? Can anybody share protocols of doing it? I am trying to uderstand this article http://www.sciencedirect.com/science/article/pii/S0890850804001008 , 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.

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.

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

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