(by tk from shaggy.csail.mit.edu)
Tue Oct 16 21:18:20 EST 2007
"Solaiman, Dan" <Dan.Solaiman from ARS.USDA.GOV> writes:
> I would be most interested in learning more about this. I had past
> experience of problem read-through when there is a hairpin structure.
> DMSO or betaine did not help me. I had to manually read the weak
> sequencing peaks of the ABI output to barely make out the sequence. If
> other people have better solution I would be grateful to learn more
> about it.
You could also try raising the extension temperature of the sequencing
reaction from 60 to 66 or 68 degrees. This would tend to destabilize
hairpins. A cycle from 60 to 70 briefly and back to 60 again might
also work well.
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