Sequencing close to the primer

David F.Bishop david.bishop at mssm.edu
Thu Feb 1 09:51:02 EST 2001

I agree, if one uses a spin column to remove dye terminators, one can 
read accurately from the first nucleotide.  If you use short spin 
columns, as in 96-well plates, you may have to do some optimization 
or evaluation of vendors to find the conditions and manufacturer that 
gives consistently clean results.

-David Bishop

>The new dGTP BigDye kit should have very good signal at the beginning of your
>Our experience is, if you have a very good and clean template, even using
>dRhodamine or BigBye can read right from the first nucleotide after the primer
>DNA Sequencing Facility wrote:
>>  I have a customer who would like to be able to read within 15 bases of the
>>  end of the primer.  I have a 373 ABI sequencer.  It seems that there is
>>  some variety in how close to the primer the sequencer will read.  Any
>>  suggestions?
>>  Thanks,
>>  Paula
>>  ---
>Department of Ophthalmology and Visual Sciences
>Chinese University of Hong Kong
>3/F Hong Kong Eye Hospital
>147K Argyle Street, Hong Kong
>Tel: (852) 27623152 Fax: (852) 27159490
>email: leungyukfai at hotmail.com, yfleung at cuhk.edu.hk
>URL: http://i.am/yfleung (My functional genomics home)

David F. Bishop, Ph.D.              | EMail: david.bishop at mssm.edu
Professor of Human Genetics         | Phone: (212) 659-6795
Dept. of Human Genetics, Box 1498   | FAX:   (212) 849-2508
Mount Sinai School of Medicine      |
1425 Madison Avenue, NY, NY 10029   | www.mssm.edu/genetics/home-page.html


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