jb031504 at mbcr.bcm.tmc.edu (Jim Bone) writes:
>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.
What also works is to scale down the standard protocol by 50% and use a
Qiagen midi column. I found you get almost as much DNA that way.