Research Genetics MapPairs PCR primers

juleen.cavanaugh at anu.edu.au juleen.cavanaugh at anu.edu.au
Tue Jun 11 23:12:15 EST 1996


In <wolffie-1006961029300001 at 128.95.13.235>, wolffie at u.washington.edu (john wolff) writes:
>We frequently get "dud" MapPairs primer sets from Research Genetics.
>
>We use a lot of different markers and we spend a lot of time trying to
>optimize conditions to coax the "duds" into working, with spotty success.
>
>My question:  is the problem with the RG MapPairs, or am I just too plain
>stupid to use them??
>
>When I get a dud, I try varying the Mg++, the annealing temp, and
>sometimes try 10% glycerol.  We use an MJ cycler PCT-100 with a 92C
>melting temp.  Mostly these are Weissenbach primers, whose conditions may
>never have been too carefully optimized to begin with.  Often, I give up.
>
I also spend time optimizing the mappairs -its worth it, but check out the 
optimization listing on GDB - we have had some Res Genetics primers that 
wouldnt give the correct size product no matter what we did - then checked 
GDB and that  amplification required weird (I mean weird!!!!) conditions - presto
- correct product!  Other mappairs are definitely dud.

Juleen Cavanaugh



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