Mark D. Garfinkel
mg16 at ellis.uchicago.edu
Tue Mar 15 09:51:53 EST 1994
ZACHARIAS at Meena.CC.URegina.CA (Tracey Zacharias) writes:
>I have received a cDNA library cloned into lambda nm1149 and am wondering
>what the host cells should be. [...]
I've built libraries in this vector. It's a primo lambda phage,
not like the wimpy Charon-series replacement vectors; big plaques, high
DNA yields. I loved working with it. For the benefit of readers who've
never heard of it, lambda-NM1149 is a jazzier version of gt10. By that
I mean it's a cI-insertion vector, giving the cloner a turbid-plaque vs.
clear-plaque visual screen as well as genetic selection for insert-
bearing clones. NM1149 is superior to gt10 in that it possesses a unique
EcoRI site and a unique HindIII site, both in the cI-gene allowing for
more-versatile cloning. And NM1149 has a somewhat greater maximum insert
size than gt10 (10-kb vs. 7-kb, I think). "NM" stands for Noreen Murray,
a lambdologist in Scotland, who's made several significant contributions
both to phage biology and cloning technology.
For initial plating of my library packaging mixtures, in order
to select for cI- insert-bearing clones, I used a derivative of C600
that carried the hflA150 mutation (high-frequency-lysogenization). For
subsequent screening of hybridization positives & DNA preparations from
plaque-pure positive clones, I used C600 as host. Both strains are in my
personal freezer collection; if you can't get them locally, feel free
to send me e-mail with a snail mail address & I'll send you stabs.
Mark D. Garfinkel (e-mail: garfinkl at iitmax.acc.iit.edu)
My views are my own, which is why they're copyright 1994 (c)
Ignore the header; I post from here only if I can't post from there.
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