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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