Uni-ZAP XR Mass Excision Jam

Reinhard Eckloff reckloff at hortresearch.co.nz
Tue Mar 27 17:59:24 EST 2001


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
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Private Bag 92 169
Auckland
New Zealand
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