Re: BAC preps
Robert Lyons (boblyons@umich.edu)
Thu, 07 Oct 1999 08:41:40 -0400
Deb Grove wrote:
>
> Hi
> We have done 90K BACs (700 to 800 base read) and more recently
> 190K BACs with about 500 base read. We also do quite a few smaller cosmids
> and BACs as well. These are prepared by several different labs on campus
> and we have students, post-docs, and techs preparing them.
> Qiagen and spec quantitation were the primary methods. The 190K was
> purified using Clonetech-Nucleobond plasmid kit -- midi size. Quantified
> with spec but a gel was also run to verify the amount. Clonetech is
> interested in using the electropherogram in a promo flyer.
To Deb's comments, I would only add one note. We also routinely sequence
BACs and other large-insert clones. Qiagen preps seem to be the best bet
for reliable reads (ca. 700). If a prep fails, however, we've found that
for some reason an extra phenol-chloroform extraction can convert a "bad"
prep to a good one. Make sure you carefully remove the phenol and chloro-
form after the extraction, as always.
Bob Lyons
-----------------------------
Robert Lyons, Ph.D.
Director, DNA Sequencing Core
University of Michigan
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