Does Taq have reverse transcriptase activity? - (Aug/31/2005 )
QUOTE (Marvilla @ Aug 31 2005, 06:33 PM)
I've perform a reverse transcription control without reverse transcriptase to see if my sample contains genomic DNA. I previously digested RNA with DNase I RNase free and I made sure of completness of DNA digestion. Then I perform PCR of b-actin and I got amplification even in the RT minus control. So I think that if Taq has got reverse transcriptase activity then RNA template for reverse trancription is being amplificated in the subsequently PCR. what do you think?
I met a similar problem with Qiagen hotstart Taq. In RT- control, one gene showed both 'genomic' band and 'RNA' band before DNase I digestion. After DNase I digestion, the 'genomic' band disappeared but the 'RNA' band still existed. My 'water' negative control showed no band. Have you eventually got an answer about the reverse transcriptase acivity of Taq?
ps answers or comments from anybody else is welcome.