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
