BAC Libraries -- How To?????

Shaun Tyler styler at mb.sympatico.ca
Sun Feb 20 19:08:31 EST 2000


I am about to attempt constructing a BAC library and I’m looking for
some technical advice.  My primary query at this point has to do with
preparation of the inserts but since I’ve never made this type of
library before anything you might have to offer about the process would
be appreciated.

What is the best way to obtain the inserts?  I assume the partials will
have to be recovered from a PFGE gel and I’m not quite sure what
conditions to use for running the gel.  What is the best size range to
select and what conditions should be used to minimize the distribution
of these fragments over the gel?  I’ve done a reasonable amount of PFGE
in the past but usually the goal has been to spread the fragments out as
much as possible rather than trying to compact a particular size range.
I doubt I’d have much success if I was trying to recover the partials
from several ml’s of gel but then again maybe this is how it’s done????
I haven’t been able to find much in the way of documented procedures for
this so even suggestions on where to find detailed protocols would be
appreciated.

Shaun

Shaun Tyler
Head, DNA Core Facility
Canadian Science Centre for Human and Animal Health
1015 Arlington St., Suite H3130
Winnipeg, MB   R3E 3R2
Ph:      204-789-6030
Fax:    204-789-2018
EMail:    shaun_tyler at hc-sc.gc.ca




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