Contact: Jun Fan PhD 304-696-7358
We provide dye terminator sequencing for plasmid and PCR templates using the ABI 3130 Genetic Analyzer with the capacity to sequence 96 templates per day. 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 online 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).
|PCR Product Size||Amount(ng)|
For double stranded DNA:
pmoles/ul = <span>(concentration of DNA in ug/ul)(106)
(# of bases)(650)
Primer Requirements: Please send 50 pmoles of primer per sequencing reaction either as a dried pellet or as a 10 pmole per microliter solution in deionized water.
Price List: Dye terminator sequencing:
Marshall University: $10.00 per template
All others: $15.00 per template
SNP/STR Genotyping: please inquire
Sequence Data and Analysis: You will be given sequence data generated by automated base calling software in the form of text files (.txt) and standard curve files (.ab1.) Txt files can be read with standard word processing software such as Notepad. .ab1 and .scf files can be read by Chromas software which can be downloaded from http://www.technelysium.com.au/chromas.html. We provide access to Invitrogen Vector NTI software for sequence alignment and other forms of analysis.