DNA Seq

susan fetics (sfetics@server.nybc.org)
Wed, 16 Jun 1999 10:27:41 -0400

hi-
i'm new at dna sequencing. i use the PE 373XL, 64 lanes. i am getting no
sequences from PCR samples. my protocol for all samples is: PCR 25 cycles
in 4uL halfterm (Genpak) and 2uL BigDye (ABI), centrifuge thru priceton sep
columns, dry the samples, denature samples at 90 degrees C for 3 minutes in
5 uL EDTA and deionized formamide, then i load the samples in the comb.
should i be treating the pcr samples differnt than the other template
samples? my other samples (DNA) and all the controls work fine, except for
a few misses. but i am getting blank lanes any time i try to sequence pcr
samples.

thanks!

-susan
___________________________________________________________________

Susan K Fetics
Laboratory of MicroChemistry
Lindsley F. Kimball Research Institute
The New York Blood Center
310 E. 67th St., 3rd floor
New York, NY 10021

phone: (212) 570-3188
sfetics@nybc.org