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DNA prep for P1 clones? Qiagen or PEG?

Sea Otter jsinger at nyx.cs.du.edu
Wed May 18 00:19:54 EST 1994

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.

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