Endonuclease activity: Crude extraction from E.coli

Julian Wood woodj at acs.ucalgary.ca
Tue Aug 1 14:34:07 EST 1995


I'm working on a restriction/methylation system in Rhizobium. A construct
where the methylase has been knocked down and the endonuclease is intact,
has been introduced into DH10B cells. The culture is growing slowly and
many cell wall debris are present in the culture. This is in favour of an
active endonuclease.

I'm looking for a quick method to make a crude extraction containing the
endonuclease (and possibly get rid of non-specific endonuclease activity).

The extract will be used to test for the activity of the endonuclease on a
target DNA.

Thanks in advance for any help

Patrice Rochepeau, PhD            e-mail: prochepe at acs2.acs.ucalgary.ca
University of Calgary
2500 University Dr.N.W.
Calgary, AB, T2N 1N4
Canada



More information about the Methods mailing list