Automatic sequence analyzer???

Takbum Ohn ahnss at hitel.kol.co.kr
Thu Dec 5 07:30:50 EST 1996


Hello.. netters..

I am performing sequencing by automatic sequence analyzer(ALFexpress,
Pharmarcia) for 1.1 kb gene (GC rich, about 70%) located in nearby
universal primer.
At first trial using universal primer it read through the sequence about
660 bp in length, but from site of 350 bp some errors were shown. so, i
made a 19mer primer at the site about 300 bps for directional sequencing.
In this time, there are no peaks except only the primer. The operator won't
read the sequence. At first, i just doubt the purity the plasmid sample, so
i used colume for the plasmid DNA purity, desperately, the result was the
same. Why??

I used the template plasmid in 5-10 micrograms and 100 pmol of the primer.

Any help or suggestion would be greatly aprepriated.

Tak-bum Ohn.

E. mail ad. ahnss at hitel.kol.co.kr



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