ABI kit Genotyping PCR help

radomski radomski at interchange.ubc.ca
Tue Sep 4 16:29:51 EST 2001


While carrying out genotyping using ABI's LMS-MD10 kit using MJ Tetrads
for the thermocycling, we have noticed some data quality/failure rate
issues that we think might be related to using 96-well PCR heads versus
384-well heads.

For the 96-well format we use 5 degree offset lid tracking. And for
384-well we use 80 degree constant. For the thermocycling parameters, we
use the standard ABI protocol.

Before changing over to 384-well format, we spoke to MJ tech. support and
they said that the only parameter we should have to change is the lid
setting.

Now with our current problems, and a subsequent call to tech. support,
they suggested decreasing the denaturation temperature. 

So, before starting to optimized this and other parameters, does anyone
have any hints to offer?

Thanks!

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