RT-PCR

John Watson ajwatson at romeo.caltech.edu
Thu Apr 28 11:20:26 EST 1994


In article <-260494103555 at dent-226-97.dentistry.umich.edu>, @umich.edu
(Steven Coon) wrote:
> 
> Hi netters,
> 
>     I am about to reverse transcribe the RNA and was considering using
> Clontech's 1st strand cDNA synthesis kit designed for this purpose. I would
> like any feedback on the kit or suggestions for alternatives.
> 
> thanx
> 
> Steve Coon
> sdcoon at umich.edu
> University of Michigan Medical Center


I have used Clontech 1st strand kits as well as BRl kits and home-made
reaction
mixes.  They all seem to work just fine.  I kind of like the Clontech kit
over the BRL kit.

I have not found thast DNasing the sample is necessary, but we were PCRing
over
a large intron and so would not expect DNA contamination.  I guess it all
comes
down to how much you know about your expected product.  The -RT control,
mentioned in a previous response is a good idea, IMHO.

John



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