PCR product purifn. for sequencing

Shiao Y. Wang sywang at whale.st.usm.edu
Wed Jul 6 23:33:44 EST 1994


Scrimshaw, Brian J (SCRIMSHAWB%GRIEHFS at LINCOLN.CRI.NZ) wrote:
: nice ladders. What cycle sequencing system do you use? I'd be
: interested to know how you find manual vs cycle sequencing for
: PCR products.

I have been using the cycle sequencing kit from Epicentre called
SequiTherm. Not bad. Signal pretty faint with 35S after 2 day exposure.
Can read 300 bases usually. I don't think the problem is with the kit
but with the amount of template DNA I start out with. I need to optimize my
recovery of template DNA prior to sequencing. I'm using one biotinylated
primer and recovering the ssDNA with magnetic strepavidin beads.

Shiao Wang
University of Southern Mississippi




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