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Incorporate biotinylated-dNTPs by a polymerase - biotin (Apr/14/2009 )

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 <_<

-sisi-

sisi on Apr 15 2009, 06:37 AM said:

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.

-klinmed-