DNA isolation SOP

Kajetan Groicher groi9663 at uidaho.edu
Tue Aug 18 16:36:21 EST 1998


Pim-

So far as S. aureus the standard alkaline lysis method will work just
fine with a slight modification:

	After resuspending the washed pellet in resuspension buffer, add 10ul
of 10mg/ml lysostaphin, incubate at 37C until the solution is clear and
viscous.  Incubation times range from 30min to 1hr.

The S. aureus cell wall is very tough, elimination of this step will
result in VERY poor yields, I have not tried to use lysozyme.

The remaining steps can be done as per std protocol.  For a standard
protocol try the Current Protocols or Maniatis (Molecular Cloning)
References.

I have also used this technique with great success with the promega and
quiagen mini prep kits (no affiliation).

For E. coli, any standard technique has worked fine for me, Pseudomonas
and B. sub i have no experience with.
Cheers-
Kajetan



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