Sanger Sequencing




Automated DNA sequencing is accomplished with two ABI Model 3130XL Genetic Analyzers with four-color fluorescence-based sequencing.

Contact Tom Cunningham.

Applications Single and double stranded genomic DNA, Plasmids, PCR product
Abilities Base calls accurately up to ~700 bp or greater
Disadvantages Quality of sequencing greatly affected by primer concentrations, GC content, quality and quantity of DNA
Primers offered by MRC M13F(-21), M13R(-48 or -24), T7terminator, T7, BGH rev, SP6, T3
Plasmid sample amounts required
(ngs) in nuclease-free water
sample volume 12 µl in 0.5 ml tube
Single Stranded DNA 10-50 ng
Plasmid(s) 700 ng
Cosmid or Bac DNA Contact us
Genomic DNA Contact us
PCR sample amounts required (ngs) in nuclease-free water sample volume 12 µl in 0.5 ml tube 100 to 200 bp 3-10 ng
200-500 bp 10-20 ng
500 to 1000 bp 20-40 ng
1000-2000 bp 40-50 ng
>2000 bp >50 ng
Primer concentration required 10 pmol
TempliPhi requirements 50 µl of fresh (19 hr incubation) plasmid bacterial culture
Software for analysis ABI Sequence Scanning Software
Chromas lite
Sequencher
Cost UT: $5
Non-UT academic: $6
Non-academic: $7
Complex/difficult samples academic: $12
Complex/difficult non-academic: $14
TempliPhi: $2.50
Special Instructions All samples are submitted through DNAlims. To set up or access your account, visit DNA Core Lims.pdf
After you submit your samples please bring a copy of the submission form with your samples; clearly label tubes with initials and sample number.