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!
---
