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Problems with acid phenol treated plasmid for sequencing

Song-Kun Shyue shsu at HEART.MED.UTH.TMC.EDU
Wed Dec 27 20:02:47 EST 1995


Dear net users:

        Does anyone sequenced the DNA after acid phenol (pH < 4.0) extraction?
I got very low or no signal from those plasmid by using Sequenase protocol.
I wonder that would be the problem of denaturing, so, I used NaOH to final
from 0.7 N to 1 N and still the same, other plasmids work fine with the same
protocol. Does anyone knows what happen to those DNA which can be enzyme
digested well. And the transfection to COS1 cells are good too.

Thank you in advance!

Song




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