[Q] ThermoSequenase aging
kofoid at biology.utah.edu
Thu Jun 20 12:30:34 EST 1996
I began using Amersham's ThermoSequenase kit in February, and got
excellent results with double stranded PCR templates. After a couple of
months, I began to notice some banding in the sequence ladders, but the
reagents were nearly exhausted so I simply bought a new kit.
Results were good at first but massive banding began occuring after only a
couple of weeks. All reagents from both kits were stored at -20 C or kept
on ice when in use. I use end-labelled primers and have purchased new
label and polynucleotide kinase with no improvement. I have double checked
primer/template pairs which worked well the first time and find heavy
banding with the aged kits.
Before I start the lengthy task of checking each reagent, can anybody
offer any suggestions? Is this a particularly fragile enzyme? Do the
termination mixes age unusually fast? Has anybody else noticed a short
shelf life for this kit?
Eric Kofoid Dept.Biology, U.Utah kofoid at biology.utah.edu
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