Does anyone have experience of using CA repeat primers with an anchoring
base on the 3prime end to sequence outwards from GT/CA microsatellite
repeats in cosmid clones (to give one of the flanking sequences)
using AmpliTaq FS. We can get it to work on some clones but at the
moment its pretty unreliable. It seems to work better using
radioactive methods than fluorescent. Has anyone tried to systematically
optimise the annealing temperature for these primers. We've tried both
(CA)7N and (CA)10N primers without too much success.
Nigel Holmes
Animal Health Trust
PO Box 5
Newmarket
Suffolk
CB8 7DW
UK
