Robert Hartley rh at mblab.gla.ac.uk
Thu Aug 24 03:50:18 EST 2000

In article <39A42865.E49C7F7E at po.cwru.edu>, Tim Spahlinger
<txs at po.cwru.edu> wrote:

> Hi,
>     I'm inviting your comments about commercial RT-PCR kits.  I haven't
> had any experience with the several kits on the market, but the
> so-called "one-step" kits look very appealing (looking at the company's
> literature).  I'm interested in extracting viral RNA from plasma, doing
> the RT-PCR, and then doing standard PCR to yield cDNA.  I expect the
> amount of RNA that will be extracted will be relatively low.
>     If you would like to share your experience with a kit or kit(s),
> please reply to this request.
>     THANKS in advance for your help!!!!

The thing that is against kits is you are curtailing yourself to the same
manufacturers RT and PCR enzymes.

Why not do it the cheap way and use Superscript 2 (life tech) and then
some Taq or other enzyme from another company (or even life tech). I use
SS2 and some of DNamp enzymes from Duncan (well from his company). I've
never had any hastle with it and have found him only a phone call or
posting/e-mail away.

I used to always use kits in the begining and a lot of these one step
(since I was teaching myself molbiol from a book) but now find that I am
now going back to the home made solutions and enzyme only mixes. Even for
things like RNA isolation I now use a home made lysis and then extract.  

Same for RT-PCR. SS2 and DNamp stuff. ( I think it is called Calypso).

I must also say that I have no affiliaton with Duncan or his company.

Bob; Bright (but cloudy) Scotland (again)

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