direct sequencing of long DNA fragments on ABI 377

George S.Grills grills at aecom.yu.edu
Tue May 15 10:30:45 EST 2001


Hi Krzysztof,

We have developed a protocol on the ABI 377 to get at least a 800 bases at
100% accuracy and 1000 bases at 98% accuracy using Long Ranger gels and
porous combs.  I will send the protocol to you.  I would be happy to send
this protocol to anyone else that is interested. 

- George



At 02:09 PM 5/14/01 +0100, Krzysztof Lubieniecki wrote:
>Hello everyone,
>
>I want to sequnce PCR product of about 1000 bp according to Applied
>Biosystems' user bulletin (subject: Achieving Longer High-Accuracy
>Reads on the 377 Sequencer - updated version 03/2001). In the bulletin
>it is recommended to use 4.25% PAGE-PLUS gel. I am wondering if anyone
>tried using Long Ranger instead of the recommended PAGE-PLUS
>gel? If so what were your results and what percentage of gel did you
>use?
>Also, does anyone have another method for sequencing long DNA
>fragments?
>
>Thanks in advance,
>
>Krzysztof Lubieniecki
>Immunogenetics Department
>Institute of Genetics and Animal Breeding
>Warsaw
>
>
>---
>

________________________________________________________________

 George Grills
 Director, DNA Sequencing, Genotyping and GeneChip Microarray Facilities
 Director, DNA Sequencing Group, Einstein Genome Center
 Albert Einstein College of Medicine
 713 Ullmann Building           
 1300 Morris Park Avenue       
 Bronx, New York 10461-1602    
 
 Tel: (718) 430-2657
 Fax: (718) 430-8778
 E-mail: grills at aecom.yu.edu
 DNA Sequencing: http://leper1.ca.aecom.yu.edu/dnacore
 Genome Center: http://sequence.aecom.yu.edu/chr12
________________________________________________________________


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