andrew.bettany at bbsrc.ac.uk
Thu Dec 12 07:50:43 EST 2002
We have been routinely using a RedTaq for screening purposes (i.e. PCR
amplification of a single (known) band) and have had no problems at all with
it. However, it may be because we started the whole procedure with RedTaq
and so got our PCR conditions worked out for (as mentioned in the other two
replies). Also, in our case we are using a complete mix (though a lot less
than suggested) so perhaps you would expect it to work!
Have you tried using a ready-optimised 2x mix? In our experience you can use
6ul (not the 25ul the manufacturers state) and it works nicely.
Don't know if this helps at all?
Dr. Andy Bettany
Institute of Grassland and Environmental Research
"Gerd Nilsen" <gerdn at ibg.uit.no> wrote in message
news:3df75806.28670734 at news.uit.no...
> I have recieved free samples of kits using REDTaq, at several
> occations. Sounds like a good idea every time, but it never works.
> Also other pople in the lab have tried REDTaq and gotten the same
> depressing result.
> Still, the thing has survived in the market for many years, so there
> must be some ways of getting it to work:
> So, what's the trick?
> My last trial was with the REDExtract-N-Amp Plant PCR Kit. This is a
> kit for DNA isolation and PCR.
> To troubleshoot, I skiped the plant material and just added clean DNA
> and primers to the PCR kit. plant DNA (6 ng), 10-mer primer, 95 C
> denaturing, 35 C annealing (don't ask me why, I just stept into a
> ongoing procedure to test the new kit), 72 C elongation, 40 cycles.
> Works like a charm with Dynazyme, but not with the REDExtract-N-Amp
> Plant PCR Kit.
> Any idéa?
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