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Old 03-19-2018, 06:48 AM   #7
Senior Member
Location: CT

Join Date: Apr 2015
Posts: 243

thermal issue is certainly on my mind since these are custom amplicons (16S v4). I've asked the engineer to bring a thermocouple to test it. 2 years ago the second machine (the one that's currently working) would pass the thermal ramp test but clearly not hold the correct temperature when we checked with an external thermocouple so had to have it's temp control replaced.

FWIW I use sequencing primers with locking nucleotides to increase the TM since the common v4 primers (Caporasso/Kozich primers) have TM ~62-65
Microbial ecologist, running a sequencing core. I have lots of strong opinions on how to survey communities, pretty sure some are even correct.
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