ABI 377 Hints.

Adrian Jenkins ajenkins at nibsc.ac.uk
Wed Oct 25 02:55:14 EST 2000



Hi all,

I was wondering whether any one could help me.

I have been using a 377  now for just over a year on PCR products with out
any major problems, though my average sequence reads are about 500-600 base
pairs using a 3.5 hour run time.  The machine belongs to another division,
and the person who was responsible for it and had the expertise has
recently left.

I have been using peg precipitation of the PCr product and resuspending in
dd water.  However I have recently found that this method is not suitable
for PCR products < 400bp

However, my colleagues occasionally submit samples to me to sequence, both
PCR and midi/maxi preps purified by an assortment of methods.

Reading the posts about the 3700, there are obviously hints or guidelines
to follow for automated sequencing such as resuspend your DNA in water
rather than the Buffers suggested by the kit.

Anyway the question is "Are there sites which have general rules/guidlines
for the preparation of DNA and for running the samples?" 


I have found some which suggest and discuss purification methods but there
might be some additional useful sites out there.

I welcome any comments, and if I get enough suggestions, I shall be happy
to post a summary


Adrian
-- 

Enjoy....

Adrian


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        NATIONAL INSTITUTE FOR BIOLOGICAL STANDARDS AND CONTROL

        Adrian Jenkins          Molecular Virology Group
                                Division of Retrovirology               
                                e-mail     ajenkins at nibsc.ac.uk         
                                http://www.nibsc.ac.uk/      
          
        "No plan survives contact with the enemy"  von Moltke

        "It's not the rather fast pace of life that frightens me,
	But rather the sudden stop at the end."
                                                          Anon


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