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
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-