We provide dye terminator sequencing for plasmid, PCR and BAC templates using the ABI 3130 Genetic Analyzer.  In dye terminator sequencing, DNA fragments are labeled by incorporation of a dye labeled dideoxynucleotide.  Investigators must provide unlabeled primers and purified template DNA.  Our read lengths average 800 bp for column purified plasmid DNA or PCR products. Please complete our sequencing request form prior to mailing your template DNA and primer.

Template DNA requirements:  For plasmid DNA, you should send 0.5 - 1 ug of DNA and provide the size of the insert (if known) on the request form.  We recommend use of Qiagen or Promega kits for plasmid purification and Centricon-100 filters for PCR product purification.  The amount of PCR template needed depends on the size of the PCR product (as shown in the table below).        

For double stranded DNA:

pmoles/ul = (concentration of DNA in ug/ul)(10⁶)
                                             (# of bases)(650)

PRICE LIST

Dye terminator sequencing:

            Marshall University:               $10.00 per template

            All others:                               $15.00 per template

SNP/STR Genotyping:                     please inquire

Sequence Data: You will be given sequence data generated by automated base calling software in the form of text files (.seq) and standard curve files (.scf.) .seq files can be read with standard word processing software such as notepad; scf files can be read by Chromas software which can be downloaded from: . http://www.technelysium.com.au/chromas.html We do not manually edit sequences

Shipping:  If you are sending your own primers for dye terminator sequencing, you may ship primers in solution (deionized water) or as dried pellets.  Please send your templates as ethanol precipitated pellets by next day mail service (e.g. FEDEX or UPS) to:

Donald A. Primerano, PhD
Marshall University Genomics Core Facility
Room 120 Medical Education Building
1542 Spring Valley Drive
Huntington, WV 25704-9388