About Compressions with Sequencing

daj (David Johnston) daj at nhm.ac.uk
Mon Jun 27 02:24:24 EST 1994


>On 24 Jun 1994 13:55:05 -0700,
>  Darrick Carter writes:
>
>
>
>Does anyone have a good suggestion about how to sequence a very G-C rich
>genome? We've tried formamide in the gel and deaza-guanine, but even
>these don't seem to get rid of the compressions. Any help?
>
For T7 sequencing, try dITP as supplied in the Sequenase kits and have 
DMSO at 10% in all stages. You should run a standard dNTP reaction set 
alongside as the dITP sequence, though it resolves compressions well, 
doesn't give a particularly neat ladder (more stops than normal).

Hope this helps,
DAJ

David A. Johnston
Dept of Zoology, The Natural History Museum, Cromwell Road,
South Kensington, London SW7 5DB. England
(tel 071 9389297, fax 071 9388754, email daj at nhm.ac.uk)



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