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Sequencing AT rich Templates

Mick Jones m.d.jones at imperial.ac.uk
Tue Nov 11 11:31:19 EST 2003


Hi

What is the best method to sequence AT-rich templates on a 3700 capillary
sequencer.

We have been given samples to sequence that contain ~800+ bp inserts in a
standard pUC based plasmid.  The inserts have been generated by PCR
amplification of bisulphite treated DNA (Cs gone to Us, 5meC remain as C) so
they are very AT-rich.

We seem to get good data on some templates, on others we get slippage.  Also
the signal strength drops rapidly.

Any advice gratefully received.

Mick Jones

Michael D Jones, BSc, PhD, ILTM
Head Genomics Core Laboratory
MRC Clinical Sciences Centre
Hammersmith Hospital Campus
London, UK





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