andrew.bettany at bbsrc.ac.uk
Mon Dec 16 04:05:51 EST 2002
Sorry if I wasn't clear.
We use the kit as supplied, but the manufacturer's suggest a 50ul PCR
reaction (i.e. 25ul 2xRedTaq mix + 25ul DNA + primers). Since we don't have
an enormous budget, we tried scaling this down. We routinely use 6ul
2xRedTaq mix and 6ul DNA + primers (250nM - we could probably reduce that
too, but we have _never_ run out of primers!) and it works fine for us. You
might be able to go below that, but we use 5ul cDNA in our reactions, so
there is no scope for us to do so.
"Gerd Nilsen" <gerdn at ibg.uit.no> wrote in message
news:3dfcb53d.53817828 at news.uit.no...
> 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?
> > >lots of bits snipped> > >
More information about the Methods