Shared Facilities Research

Molecular Genetics & DNA Sequencing Core Lab

Scientific Director:  Bruce Stanley, PhD   

The Core provides full service automated DNA sequencing, as well as multiplexed SNP and Genotyping analyses (SNPlex Analyses). An NSF grant funded the initial major equipment in this laboratory.

Individual investigators provide the purified plasmid or PCR template as well as the primer, unless a standard primer (T3, T7, M13, etc.) stocked by the Core Facility is requested. The sequencing reactions are performed by cycle sequencing using fluorescent dye-labeled dideoxy terminators (ddNTPs) and a modified Taq polymerase. Resolution of the sequencing products is achieved by running the spin column-purified samples on a 48-cm acrylamide gel, which gives better resolution than the 36-cm gels used in some facilities, or on a newer ABI 3130 Capillary Sequencer. Detection of the fluorescently-labeled fragments is done as the bands migrate out of the gel or capillary, using an argon laser and CCD camera.  Version 3.3 Sequencing Analysis software (using neural net tracking algorithms) is used to generate the sequencing data. Turnaround times are quick (2 days), and researchers are rapidly provided with their sequence data via email. Color-print electropherograms showing the fluorescence peak traces that represent the actual dye-labeled fragments as they leave the gel or capillary are also provided.

Services Offered:

·         DNA Sequencing from plasmids, PCR products, and BACs

·         SNP Analysis (Quantitative and SNPlex analyses)

·         Genotyping Analysis

·         Sequence analysis programs SEQWeb and the GCG/Wisconsin Package

Major Equipment:

·         ABI 3130 Capillary Sequencer

·         Applied Biosystems 377 Gel-based Sequencer