ssDNA (M13) extract. with silica?
kraev at bc.biol.ethz.ch
Tue Feb 11 06:47:06 EST 1997
<Pascal.Mertens-1002971653490001 at immuno-wawa.sciences.fundp.ac.be>,
Pascal.Mertens at fundp.ac.be (Pascal Mertens) wrote:
> I wonder if it's possible to use this silica milk to purify M13 ssDNA
> directly from culture supernatant (or from PEG-precipitated virions) for
> sequencing in place of the usual phenol-chloroform extraction?
> Does somebody have a nice protocol for that? (or any idea how to proceed).
> Many thanks in advance
M13 DNA for sequencing can be "extracted" by running the Sequenase
protocol (e.g. NaOH, neutralization etc) on the resuspended PEG pellet
directly. Alternatively, check the Sanger Centre protocol page for the
Thermomax protocol, which basically involves
heating phage pellet in Triton X100 and using it in cycle sequencing. The
latter also works
well for fluorescent sequencing.
Alexander Kraev, PhD
Biochemie III, ETHZ Zurich
e-mail kraev at bc.biol.ethz.ch
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