DNA Sequencing

DNA Sequencing Services

• ABI 3130xl Genetic Analyzer
• ABI BigDye v3.1 Terminator chemistry
• Sequence multiple sample types
  • BAC DNA
  • ds plasmid DNA
  • PCR product
• Results provided in multiple formats (.seq and .ab1)
• Rapid turn-around time (1-3 business days)
• High volume discounts

DNA Sequencing Service Fees

*Please note, samples are charged in multiples of 8.

**Project and optimization rates may be available.  Please contact FADSS for more details.

The user may request a rerun of samples with poor results. If subsequent results do not improve, the user will be charged the <1/2 plate DNA sequencing rate. If subsequent results improve, the user will not be charged for the reruns.

Template Considerations

Of primary importance to the success of sequencing reactions is the quality and quantity of sample provided. FADSS cannot be held responsible for failures in sequencing which are related to poor preparation of the DNA template used, or failure to follow recommended guidelines.

• Contaminated template will give high background noise and poor, or no sequence information
• We recommend many of the commercially-available extraction and purification kits (eg. Invitrogen, Qiagen, Promega, Wizard Plus, Sigma)
• It is critical that the template and primer concentrations are measured carefully, as incorrect quantitation (whether higher or lower) will result in poor sequencing results.
• Template must be re-suspended in water or Tris, not TE buffer, as EDTA will interfere with the ion concentration in the sequencing reaction

Primer Considerations

When designing custom sequencing primers, we recommend:

• T_m between 55˚C and 60˚C
• “G” or “C” at 3’ end
• Avoid primers with runs of 3 or more of the same base
• Use primer design software to avoid primers with hairpin or dimmer characteristics
• Primers must be re-suspended in water or Tris, not TE buffer, as EDTA will interfere with the ion concentration in the sequencing reaction

Sample Submission Guidelines

1. Please provide 15ul of template DNA and primer at the appropriate concentration (refer to table below) in water or Tris (not TE!)

2. Please submit samples in 8-well strip tubes or in plates.  For plates being shipped to FADSS, we recommend chimney well plates (Fisher catalog # 8408230) to minimize well-well contamination and evaporation.  Place clearly labeled samples on the top shelf of the freezer in SCI355. The plate and or rack label should match the Plate/Rack ID.  If you are shipping your samples to FADSS, please send your samples to the address at the top of the DNA Sequencing Request form Attn: FADSS.

3. Please submit the completed Sequencing Request Form to fadss.ok@ubc.ca. Incomplete request forms may result in delayed processing.

Typical turnaround time is 1-3 business days upon receipt of sample and request form submission. Please allow more time for large submissions (multiple plates).

UBC O clients can obtain their sequencing results on and off campus by logging into their CWL accounts. Text files (.seq) and binary data files (.ab1) will be located in the following directory:

K:\RAA\FADSS\username

Non-UBC O clients can obtain their results at https://payforprint.ok.ubc.ca/netstorage/ using a Netstorage account and password provided by FADSS.

The .ab1 binary data files generated can be used with a variety of software programs. FADSS facilities include computers with Gene Code’s Sequencher 4.6 software. For clients who do not have access to our facilities, there are a variety of freeware programs available including:

• Finch TV
• Sequence Scanner

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Last reviewed 7/24/2012 2:35:56 PM