Host cells for Lambda FIX II?

Chris Kafer ckafer at iastate.edu
Sat Oct 17 17:02:32 EST 1998


I use P2392 with good success with my Arabidopsis and Tobacco
libraries.  The DNA is of sufficient quality and yield to
auto-flourescent sequence if purified by a simple DE52 cellulose
method.

Out of curiosity, dont you have to use a P2 strain to grow this phage?
My understanding was that only non-recombinants would grow if not.
I've been unable to find a decent explanation of the P2 selection in
our books here in lab though...






On 16 Oct 1998 18:18:28 -0700, dhavilan at IMM2.IMM.UTH.TMC.EDU ("David
L. Haviland, PhD") wrote:

>On Fri, 16 Oct 1998, Scott Henderson wrote:
>
>> What host cells work well to product high titer stocks of Lambda FIX II
>> besides XL1-Blue MRA and XL1-Blue MRA(P2)?  I've tried XL1-Blue MRA(P2)
>> without success.  The problem could be my own stock of cells but I am
>> hesitant to buy more of the same and waste time if something else will work
>> as well or better.
>
>Scott:
>
>I'm in that boat too, but I looked around for different reasons.  What I
>don't like about the XL-1 MRA's is that there is no selection for them
>other than minimal media with thymine, no antibiotic selection.  I sat
>down and "bent" my brain a bit and recalled that the LE392 should work.
>They did, in fact we obtained great infections but the overall phage DNA
>yeild didn't compare to the XL-1 MRA, so we stuck with the MRA.  Word of
>warning, do NOT use the P2 strain for your phage minipreps.  You might get
>your desired phage DNA but it will be buried amongst a sizeable amount of
>the P2 DNA.  (been there done that error!)
>
>As I said, we had high hopes for Le392 but had to settle for the XL-1 MRA.
>
>Hope this helps,
>David
>
>==========================
>David L. Haviland, PhD
>Assistant Professor, Immunology
>University of Texas, HSC - Houston
>Institute of Molecular Medicine
>2121 W. Holcombe Blvd.
>Houston, TX  77030
>http://www.uth.tmc.edu/~dhavilan
>713.500.2413-Voice
>713.500.2424-FAX
>==========================
>
>
>




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