DNA Sequencing, Sanger Capillary

dna sequence image collageThe facility uses the 3730 DNA Analyzer from Applied Biosystems, Inc. and BigDye® Terminator Cycle Sequencing chemistry. This automated instrument utilizes fluorescent dye labeled nucleotides and provides high volume capabilities, e.g. 384 sequencing reactions in 12 hours, and typically yields 800 to 1000 bases per reaction depending upon the quality and nature of the template. Example [pdf] of good quality DNA sequence, i.e. electropherogram.

Order

For all questions concerning the template, primer, concentrations, tubes, results, etc., please see " PDF icon PDF icon Procedures For DNA Sequencing [pdf] at the Plant-Microbe Genomics Facility."

In addition, the Facility offers plasmid purification from E. coli and amplicon purification in plates with a robotic system prior to sequencing.

How to place a DNA sequencing order:

  1. Go to dnaLIMS to login (or create an account if you have not done so previously)
  2. Click on either individual or plate link under the Sequencing header
  3. Follow the instructions.

How to deliver samples

How to get results:

  1. For samples delivered to the Facility by 10:00am most days results will be available by 10:00am the next business day
  2. An email will be sent to your address when the results are available.
  3. Log into dnaLIMS and click on "Download DNA Results"

Data Analysis

In order to view, print and edit the electropherogram, i.e. ab1 file, one of the following programs is required which are free and work with Windows (W), Macintosh (M) or Linux (L).  Each program is unique with different features, and strengths.  The program Xplorer is also incorporated into dnaLIMS.

DNA Sequencing References and Resources on methodology, data interpretation and troubleshooting


[pdf] - Some links on this page are to .pdf files. These are designated by [pdf] following the link. If you require these materials in alternate format contact our web manager.

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