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|
|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
|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.