DNASeq Internal Standards

James Farmar (jfarmar@nybc.org)
Fri, 23 Jul 1999 13:20:31 -0400

We are looking for a source of pcr products as internal controls for our dna
sequencing. ABI sells a control DNA template with its Big Dye Kit but
doesn't sell an equivalent control for a pcr product. For each sequencing
run, we would like to include a DNA template, a small pcr product and a
large pcr product as controls. If correct sequence is obtained from each
control, we could avoid the usual back-and-forth between researcher and
sequencer about the cause of a failure to obtain sequence from a sample.

Our thinking is that one pcr product should be ~250 bases in size and the
other should be ~2500 bases. We can determine the best stoichiometry of Big
Dye/Half-Term chemistry for these products and then use that stoichiometry
for all pcr samples submitted for sequencing.

Ideally, these internal control pcr products should be readily sequencable
using Big Dye chemistry and should be commercially available in the
multi-microgram range.

Can anyone recommend pcr products to buy or to make?
Thank you.

Jim Farmar and Sue Fetics
Laboratory of MicroChemistry, Kimball Research Institute of the New York
Blood Center