reverse transcripatase in Taq?

Rae Nishi nospamnishir at ohsu.edu
Mon Oct 20 15:37:07 EST 1997


In article <3447D881.7FB1 at ubaclu.unibas.ch>
Peter Schuchert <schuchert at ubaclu.unibas.ch> writes:

> It was once reported that Taq can also reverse transcribe RNA.
> What's the actual opinion on that?
> 
> I try to make rt-PCR of a weakly expressed gene and get specific bands
> even without reverse transcription (contamination can be excluded,
> the RNA has been treated with DNAseI and the genomic copy has an intron,
> so that can be excluded too).
> Any hints before I go and try RNAse treatment (reluctantly!!)
> 
> cheers,
> Peter

Is this a gene you have cloned?  If you have alot of plasmid encoding
the cDNA flying around, one of your reagents could have gotten
contaminated.  Have you tried leaving out a template or testing the
water that you use in any of your reactions?  Or the oil (if you use
oil--we have even gotten contaminated oil).

reply to nishir at ohsu.edu
Rae Nishi, PhD
Professor
Dept. Cell & Developmental Biology
Oregon Health Sciences University
Portland Oregon 



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