gerdn at ibg.uit.no
Sun Dec 15 12:03:52 EST 2002
On Thu, 12 Dec 2002 12:50:43 -0000, "Andy_Bettany"
<andrew.bettany at bbsrc.ac.uk> wrote:
>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.
Now we are getting somewhere, you are modifying the procedure.
What do you mean?
6 ul total volume, or 6 ul 2 x mix in 12 ul, or ....?
Is the 2x mixture too concentrated?
>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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