The DNA Secquencing Core Facility provides:

DNA Sequencing (ABI 3730 Capillary Sequencer)

Users provide sequence-ready DNA templates. Users have a choice of adding their own sequencing primers or choosing one of the primers available through the facility (M13F, M13R, SK+, KS+, SP6, T3, and T7). The facility has posted template and primer requirements on its Intranet Web site for best results.

Accepted for sequencing are ssDNA, dsDNA, and PCR products, all of which should be prepared with respect to purity and quantity according to the recommendations listed on the facility Web site.

DNA cycle sequencing is performed using ABI Big Dye Terminator chemistry. Control samples are included in all reaction sets to assess the quality of reactions and capillary electrophoresis. Users should expect between 500 and 800 nucleotides of reliable sequence on long templates.

High-throughput sequencing (i.e., templates received in fully loaded 96 well plates) is available at a reduced cost.

Once sequencing results have been analyzed by the facility, the results are posted in the user's individual account in the form of text files and electropherograms. Turn-around times are generally between 24 and 48 hours. Sequence results are left on file for a period of one month.

Fragment Analysis (ABI 3100 Capillary Sequencer)

Users perform PCR reactions using uniform template concentrations and their own fluorescently labeled primers. Prior to analysis, samples of each PCR product are mixed with size standard provided by the facility.

Results are analyzed with Genemapper Software, printed, and made available for pickup within 2 to 4 days after drop-off.