Help: Reliable Sequencing Past 300 b.p

dmckinnon at epo.som.sunysb.edu dmckinnon at epo.som.sunysb.edu
Sat Dec 30 19:22:48 EST 1995


We are having trouble routinely producing reliable sequence data past 
approximately 300 b.p.  We use the Sequenase enzyme with 35-S-ATP on 
double-stranded plasmid DNA.  The gel system is 6% or 8% acrylamide in 
0.5x TBE.  The gel apparatus is a 50 cm BRL unit usually run at 80W for 
up to 4.5 hours.
Any suggestions on the simplest ways to improve our readable sequence 
length would be greatly appreciated.





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