PHAGE DNA KITS?

[Randall J. Cohrs]

> 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 
ETOH.

- - - 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 
different libraries.

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.

Good Luck

RJC in Denver