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DNA sequence information is of fundamental importance and is required by most research programs at St. Jude. The Hartwell Center offers High Throughput DNA Sequencing in support of of ~100 research programs and sequences 80,000 templates generating ~40 million base pairs of DNA sequence information each year. We use state of the art technology to provide you with high-throughput and high quality data.
Several DNA sequencing chemistries are available:
- Big Dye® Terminator Chemistry
- Other Sequencing Chemistry Options
- Cycle Sequencing Reaction Conditions
Currently, DNA Sequencing is routinely performed with Big Dye® Terminator (v3.1) Chemistry on Applied Biosystem 3730XL DNA Analyzers, which are capillary electrophoresis based platforms. Big Dye® Terminator Chemistry is a technique in which fluorescent dyes are incorporated into DNA extension products during cycle sequencing reactions. The incorporation of dideoxynucleotide triphosphates (dye terminators) blocks further chain elongation, resulting in truncated linear fragments labeled with one of four fluorescent dyes. After proper purification, the dye-labeled DNA extension products are robotically loaded onto the capillary array of the ABI Prism 3730XL DNA Analyzer, where the samples are electrokinetically injected into the capillaries. The labeled fragments are then separated by size during electrophoresis, which determines the order, or sequence, of the four deoxyribonucleic bases, adenine(A), cytosine(C), guanine(G), and thymine(T) that comprise the DNA molecule. Each dye, which emits light at a different wavelength, is detected by laser irradiation and automatically analyzed to determine the DNA Sequence. The DNA sequence data generated in the Hartwell Center is electronically archived and easily available via the Shared Resource Management data retrieval tool.
