PHAGE DNA KITS?
Randall J. Cohrs
COHRS-R at Oberon.HSC.Colorado.edu
Fri Oct 7 11:47:05 EST 1994
> Subject: PHAGE DNA KITS?
> Are any kits or techniques available to purify very clean DNA from
> phage preps prepared by the plate lysate method?
I've prepared phage DNA from plate lysates by PEG-precipitation
followed by twice banding on CsCl gradients.
Also by PEG precipitation without banding
but with SDS/proteinase K followed by
phenol/chloroform/isoamyl alcohol and
- - - for my money ($115 for 20 extractions) I now use Nucleobond from
the Nest group (no affiliation)
Results so far:
prep 1: from 50 ml at 1x10e7 pfu/ml yield=2.7 ug
prep 2: from 25 ml at 1x10e9 pfu/ml yield=138 ug
prep 3: from 25 ml at 1x10e9 pfu/ml yield=44 ug
prep 4: from 50 ml at 1x10e9 pfu/ml yield=117 ug
All preps are plate lysates (cDNA cloned into lambda-ZAP) from
prep 1 was stored at 4C and therefore the loss in titer.
preps 2-4 were stored at -70C in 7% DMSO, stable titer and the DMSO
doesn't seem to interfer with the extractions.
none - the - less , All DNA preps gave PCR amplifiable DNA (looking
for actin and a rare virus message)
haven't needed to compare other products.
RJC in Denver
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