Miss YY Szeto
yszeto at hgmp.mrc.ac.uk
Sun Oct 19 10:04:19 EST 1997
Our lab has tried both standard alkaine lysis and Qiagen kit for preparing
BAC DNA (from 80-200Kb). Qiagen has a protocol with modified volumes of
I handle BACs with uncut pipette tips and it turns out to be OK. But I
avoid vortexing them.
On Fri, 10 Oct 1997, Jeffrey J Meyer wrote:
> I would appreciate any suggestions from people who have worked with the
> isolation and mapping of BAC clones. We're dealing with DNA fragments on
> the order of 160kb. Which kits were used (Qiagen?) for the preps?
> (and any conditions that were changed from manual protocols (e.g.,
> volumes)) Will this size of DNA be sheared when using uncut pipet tips?
> Also, does anybody have ideal parameters for resolution of bands from
> 5-160kb on a PFGE unit? Finally, any suggestions for the production of
> extremely pure DNA (e.g., special changes to normal prep-protocols) for
> partial digestions?
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