Uni-ZAP XR Mass Excision Jam

Alejandro Miguel Martin Dunn alejandro.martin at cigb.edu.cu
Wed Mar 28 08:31:52 EST 2001


Have you tested whether the SOLR cells are still F'? Use the helper
stock to check for phage sensitivity, or grow in minimal medium without
thiamine.

cheers,
alejandro

-----Original Message-----
From: Reinhard Eckloff [mailto:reckloff at hortresearch.co.nz]
Sent: Tuesday, March 27, 2001 5:59 PM
To: methods at hgmp.mrc.ac.uk
Subject: Uni-ZAP XR Mass Excision Jam


Uni-ZAP XR Mass Excision Jam
Apparently, I have had no luck mass excising Bluescript from a Uni-ZAP
XR library. 
I have done everything according to the protocol by Stratagene. 
I have titered the library and the ExAssist helper phage before the
excision. 
I have combined 1.5 x 10E5 pfu lambda phage, 1.5 x 10E6 XL1-Blue MRF'
cells and 1.5 x 10E7 pfu ExAssist and then done as the protocol says.
But I can not get any colonies when I want to titer the phagemid with
SOLR cells on LBamp plates.

Any suggestions as to what is wrong?
The library seems OK as ESTs have been sequenced from it (and others
have excised it).
I have tested the amp plates, they are OK.
What else could I try?
Reinhard

-- 
Reinhard M G Eckloff
Hortresearch
Plant Health and Development
120 Mt Albert Rd
Private Bag 92 169
Auckland
New Zealand
Tel: +64-9-815 4200
Fax: +64-9-815 4201
Mob: +64-21-258 4720
www.hortresearch.co.nz

---




More information about the Methods mailing list