[Arabidopsis] BAC Problems

Matthew Pelletier via arab-gen%40net.bio.net (by Matthew.Pelletier from houghton.edu)
Wed Aug 15 10:32:52 EST 2007

I am in the process of trying to purify several BAC clones from the
ABRC.  Three of the clones are PAC (Mi/P1) clones and one is a TAMU
clone.  They vary in size from 16 kb inserts to about 90 kb inserts.  I
have purchased a Spin Doctor kit from Gerard Biotech that is designed to
purify BAC clones from 50 ml cultures.  My student's first attempt
yielded very little DNA.  I need to get about 20 micrograms of each of
these BAC clones.  I'm wondering if people have a sense for how large a
culture I will need to obtain this much DNA, as well as whether anyone
has a particular protocol or kit they have had success with.  Also, what
type of media do you grow these clones in?  I thought adding IPTG to the
media was supposed to increase copy number of the P1 clones.  Thanks for
any insights on this!

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