DNA sequencing services are currently provided using the Applied Biosystems 3730 DNA Analyzer. The ABI machines utilize a one-lane sequencing process with refined, four-dye fluorescent labeling methods and a real-time scanning detector. The biochemistry of one-lane sequencing is similar to Sanger-based methods. The sequencer automatically processes collected data after electrophoresis is complete. The software calls the sequence and creates files containing the analyzed data and its sequence. Because the fluorescent dyes used in one-lane sequencing are compatible with thermal cycling, the sequencing reactions can be amplified with Taq polymerase. Thermal cycling generates consistently high fluorescent signal, even from large inserts. Cycle sequencing also allows simple, accurate sequencing of double-stranded templates with secondary structure and high GC content.

The service provides standard sequencing primers for several vectors. Custom primers are provided by the user or can be synthesized in our facility. The sequencing service is presently able to handle up to 1000 samples per day and every effort is made to keep our turnaround time to a minimum.

We strongly recommend Qiagen purification of plasmid templates. PCR products should be gel isolated and purified for use in automated sequencing. Inquire at the DNA Analysis Facility for additional information about template and/or primer preparation. Please see the Protocols section for additional details on template preparation.