Denaturing ds DNA for sequencing

benedik at benedik at
Fri Mar 3 18:44:43 EST 1995

Here is a stupid question for which I am sure there is an obvious
answer that I just don't know.

When denaturing ds plasmid DNA for sequencing, why do we go to all the
trouble of adding NaOH, then neutralizing and ETOH precipitating? Why
doesn't it work just heating the DNA to 85oC then chiling on ice? I am
sure there is a reason, but I can't think of it.

Michael Benedik               benedik at
Biochemical Sciences
University of Houston
Houston, TX 77204-5934

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