Sequencing Very AT Rich sequences on a ABI 3700 with BigDye ver 2

Mick Jones mickd.jones at ntlworld.com
Sun Oct 13 02:57:20 EST 2002


We have been analysing ~900 bp inserts cloned in standard plasmid vectors,
which have been generated by bi-sulphite modification of DNA, and are thus
extremely A/T rich (in fact ~85%+) and the traces obtained with BigDye
terminator (ver. 2) on our 3700 are poor.

Has anybody any advise on sequencing such AT rich clones?

I notice on the ABI WEB site that An or Tn stretches are best sequenced with
the dRhodamine terminator kits.

Will this help solve our problem, as we don't have vast An or Tn stretches,
but just very AT sequences?

Many thanks

Mick Jones
Head Genomics Core Laboratory
MRC Clinical Sciences Centre
Imperial College, London
Email:   mick.jones at csc.mrc.ac.uk


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