Sequencing large banks

[Sandy Beeser]

  Has anyone had good experience sequencing small amounts ( say 100 bp
then its off to the database) from really large plasmids using cycle
sequencing ?  We have a Yep13 chromosomal bank ( size 17-32 kb) that I
can't get any sequence from using promega's fmol system with direct
incorporation, although it will sequence pBR322 with no problem.  Should I
just scrap direct incorporation and go for end-labelling the oligo ? 
Techincal support at promega said size should not matter, although they
said that it would require some optimisation with regards to
template/primer concentrations as well as cycling parameters.  Any help
will be appreciated.

Alexander Beeser
Grad student 
University of Tennessee, Memphis