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clean-up of ssDNA

the End jgraham at bronze.ucs.indiana.edu
Thu Aug 5 18:18:05 EST 1993

> ...isolating ssDNA....

This is a new one for me too, but I tried using Proteinase K at 70C (yes,
that's 70C) as reported in a recent Biotechniques. In that case a PEG pellet
of phage is resuspended in 300 ul of TE, and SDS added to 0.2 %, and digested 
for 15 min at "65 to 75C" with 10 ug of proteinase K. After digestion add 
40 ul of 5 M potassium acetate while hot, and transfer to ice for 10 min. 
Spin out debris and precipitate with ethanol.

Works like a charm for sequencing templates. No phenol, no sweat, no kit. :)

J. Graham
Biology and Chemistry Departments
Indaiana Unviersity Bloomington

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