What is the best method to sequence AT-rich templates on a 3700 capillary
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.
Michael D Jones, BSc, PhD, ILTM
Head Genomics Core Laboratory
MRC Clinical Sciences Centre
Hammersmith Hospital Campus