Difficulties in sequencing microsatellites

Wolfgang Latz latz at dhm.mhn.de
Tue Nov 21 08:53:30 EST 2000


Hello,
I meet some problems in sequencing a microsatellite region of about 35
CA repeats. I tried cycle sequencing using ThermoSequenase and
SequiTherm Excel II kits as well as isothermal sequencing. The first 25
repeats are clearly readable but then the polymerase seems to jump off
the template DNA and the reading stops. The template is a purified PCR
amplicon and we use a LI-COR 4000.
Has anyone an idea how to manage to sequence through that microsatellite
region. Any suggestions are welcome.

Thank you

Wolfgang






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