Incorporate biotinylated-dNTPs by a polymerase (View forum version)



sisi

Posted 14 April 2009 - 08:37 PM

Dear all,

I'm trying to use PCR to incoporate biotinylated-dNTPs into DNA, and I wonder whether anyboby here did this work before, and which is the best polymerase I should use.

Thank you so much!

Please connect if you interested in this topic!

Sisi <_<

klinmed

Posted 15 April 2009 - 09:53 AM

Dear all,

I'm trying to use PCR to incoporate biotinylated-dNTPs into DNA, and I wonder whether anyboby here did this work before, and which is the best polymerase I should use.

Thank you so much!

Please connect if you interested in this topic!

Sisi ;)

If all you want is a biotinylated product, PCR with taq as normal. Use 1 mM dATP, 1 mM dCTP, 1 mM dGTP, 0.5 mM dTTP,
0.5 mM Biotin-11-dUTP. You can alter the degree of biotin incorporation with different ratios of dTTP to Biotin-dUTP. Lower nucleotide concentrations work fine.

Hope this helps.