IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

Sequencing large banks

Sandy Beeser abeeser at utmem1.utmem.edu
Fri Oct 25 22:26:35 EST 1996


  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



More information about the Methods mailing list

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