IUBio

DNA Seq core facility set-u

Anthony Tomlinson tomlinson at pplros.demon.co.uk
Tue Oct 22 17:29:06 EST 1996


In article <53e92o$46r at net.bio.net> Lawrence Washington,
lwashing at sunflower.bio.indiana.edu writes:
>> We are in the process of setting up an automated DNA sequencing core 
>> facility here at the Salk Institute.  We will be starting out with 2 ABI 
>> 377 machines...On a good day, with 48cm 
>> plates we can get about 700 good bases maximum.  Often the reads are 
>> significantly shorter.  Therefore, one issue is how to modify the 
>> protocol to maximize read length.  Any suggestions for changes (maybe 
>> voltage, acrylamide %, running temp, buffer)?

I think it's fair to say that signal quality is the first thing to
optimise.
Especially if you are terminator sequencing, the length of read that the
software
can give you is most affected by the signal strength/quality.  We have
had a 377
for 6 months, and at first the reads were not that good, but then, as we
optimised
our template preparation and cycling conditions, the signal got better,
and
the reads were correspondingly longer.  

Anthony.




More information about the Autoseq mailing list

Send comments to us at biosci-help [At] net.bio.net