3 replies to this topic

[krildrik]

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.

[phage434]

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.

[krildrik]

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

[phage434]

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