Double-stranded DNA sequencing

"Dr." Shoumo Bhattacharya B1 061 sbhattac at
Tue Oct 22 14:04:34 EST 1991

Sandra Pena de Ortiz writes:

Hi!  Does anyone have experience with double-stranded DNA sequencing using
Sequenase version 2.o of USB?  I would appreciate ANY information for
optimization of results.  Thanks in advance.
Sandra Pena de Ortiz     : penas at ucbeh.bitnet
University of Cincinnati
College of Medicine
Toxicology Program

Use 5-10 ug DNA,  add 0.2M NaOH (final) for 30' at 37 deg.
EtOH precipitate and follow the USB protocol. Load 4ul
on the sequencing gel. DNA miniprepped by the boiling
method (Maniatis) gives good results.
Best of luck!
Shoumo Bhattacharya

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