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.