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DNA Sequencing Facility

DNA Sequencing

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Primers

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Template Prep

Sample Sheet

Instructions

Instrumentation

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Instrumentation

The OMRF DNA Sequencing facility provides DNA sequencing to all Foundation investigators. The Facility has the only ABI3730 capillary sequencers in the state of Oklahoma, capable of running 180 samples per day using single-stranded, double-stranded, or PCR amplified DNA. Operating hours are Monday through Friday, 8:30 am to 5:00 pm. For same day processing, samples should be received by 1:00 pm. Turnaround time is generally 2 days, depending on the number of users. The cost is $6 per sequence to OMRF and OUHSC investigators.

Method (from ABI Sequencing Guide)
Using Polymerase Chain Reaction, the DNA template is labeled with four different fluorescent terminal ddNTPs. The labeled DNA fragments are electrophoresed through a polymer filled capillary and separate according to length. At the end of the capillaries, they pass a laser beam. The laser light excites the fluorescent dyes, each of which emit light at a specific wavelength. The light is collected and separated according to wavelength by a spectrograph on a cooled, charge coupled device (CCD) camera. The data collection software collects the light intensities using software-selectable filters and stores them as electrical signals for eventual processing.