phage prep help really, really needed

Rae Nishi nishir at ohsu.edu
Wed Nov 9 18:37:47 EST 1994



I beat my head against the wall for quite awhile trying to isolate
phage from plate lysates and never got very good yields and also had
problems cutting the DNA.  Then I got a protocol for liquid lysate prep
and it works every time.  Just prepare an overnight 30 ml liquid lysate
culture, then isolate phage by taking sup of low speed spin and
spinning it down through a glycerol step gradient.  Treat with DNase
and Rnase, then proteinase K; chloroform phenol extract 3 times and
dialyze DNA.  Yield is typically 20-40ug of phage DNA.  If you want our
specific protocol, email me and I will send it.

Rae Nishi
Dept. Cell Biology & Anatomy
Oregon Health Sciences University
Portland Oregon 97201
nishir at ohsu.edu
**that's Orygun, NOT Ora-Gone**



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