Laurence,
You can check out our protocol for BAC end sequencing at:
http://cbil.humgen.upenn.edu:80/~atgc/physical-mapping/BAC_data/BAC_end_seq
html
We use restriction digests rather than sonicating, but either way is
probably fine
as long as the BAC is linearized.
-Eugen Buehler
ATGC
U. Penn
In article <5fps8c$69p at net.bio.net>, LHALL, LHALL at fs2.scg.man.ac.uk
writes:
> Dear all,
>> I have recently had some success at sequencing BAC'S by
> modifying cycle sequencing conditions ( ABI kit ), using more primer,
> adding DMSO etc. However, I am led to believe that sonicating BAC'S
> further improves matters. Does anyone have a protocol ?
>> P.S Does anyone have a protocol for cycle sequencing PAC'S, using ABI'S
> Diterminator kit ?
>> Regards,
>> Laurence S. Hall.
> Laurence Hall,
> School of Biological Sciences,
> 2.237 - Biomolecules,
> Division of Biochemistry,
> Stopford Building,
> University of Manchester,
> Oxford Rd.
> Manchester M13 9PT.
>>> Tel. : +44 (0)161 275 6916. E-mail :
>LHALL at fs2.scg.man.ac.uk> Fax : +44 (0)161 5082.
