PCR with TMACL

Ashok Aiyar axa12 at po.cwru.edu
Tue Nov 24 23:19:29 EST 1992


In article <2b13a0b7.1926 at lincoln.cri.nz> 
	srcksac at lincoln.cri.nz writes:

>Dear Netters,                                                                 
> I'm wondering if anyone has a reference for the use of TMACL 
>(tetramethylammonium chloride) for improving the specificity of
> PCR reactions with redundant oligonucleotides.   Thanks.
>
>     Sonya Clark.

I do not have a reference, but based on what I read on this group about year 
back, I tried a bunch of things with TMAC.  First off, it stinks terribly 
and is really most nauseating.

Secondly, and more importantly, I had to do a series of dilutions for each 
primer-pair/template combination that I used.  Ultimately, I ended up 
getting much more consistent results and increased specificity by reducing 
the final concentration of MgCl2 in the reaction, decreasing the number of 
cycles to about 15 - 18 and also decreasing the amount of primers added from 
100 pmoles / 100 ul reaction to about 15 - 20 pmoles / 100 ul reaction (i.e 
decrease the final concentration from 1 micromolar to about 0.2 micromolar).

Ashok
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  Ashok Aiyar   Department of Biochemistry, CWRU   axa12 at po.cwru.edu
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