Help for sequencing GC-rich sequence
John Lum
johshi at hotmail.com
Sat Nov 21 00:12:36 EST 1998
Hi,
I've recently cloned a promotor sequence of a gene but having big
problem in sequencing it. Only 150 ~ 200bp sequence was obtained using
the T7 npolymerase and an ALF sequencer. I suspect the problem is caused
by the GC-rich region, bcoz the sequencing result deterioate when
reaching local GC-rich region (up to 80% GC).
Switching to Cycle Sequencing using Taq polymerase doesn't help much.
Do anyone got any suggestion, kit, additives for sequencing GC-rich
DNA using automatice sequencer..??
Thanks in advances.!!
Regards,
Lum
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