DNA prep for P1 clones? Qiagen or PEG?
jb031504 at mbcr.bcm.tmc.edu
Tue May 17 17:18:21 EST 1994
In article <2ra7um$7rn at lyra.csx.cam.ac.uk>, cw117 at mole.bio.cam.ac.uk
(Charlie Wright (Genetics)) wrote:
> I'm about to do 250ml (mini in the case of P1) preps of some of the P1
> clones found on flybase. The only Protocol given says Grow O/N,
> innoculate large flask, add IPTG to induce amplification, grow some more.
> "proceed with the Qiagen maxi-prep according to the kit protocol". The
> problem is, there isn't a Qiagen for P1 bacteriophage kit. I could treat
> it like a really big plasmid, but does anyone know how the elution
> buffers should be modified (if at all?) Help, help.
> Others have suggested PEG precipitation.
> Any experience appreciated.
I just did a P1 DNA prep last week, and used a Qiagen Maxi (30 ml capacity)
column. I grew four clones, 2 did real well (>50 ug DNA), 2 did poorly. I
followed the Qiagen protocol and it turned out fine. I believe the
problems are endemic to the clones I have--they just didn't grow that well.
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