ABI 3500 Series Genetic Analyzer

Sanger sequencing targets a specific region of template DNA using an oligonucleotide sequencing primer, which binds to the DNA adjacent to the region of interest. (There must be an area of known sequence close to the target DNA.) In order to determine the sequence, Sanger sequencing makes use of chemical analogs of the four nucleotides in DNA. These analogs, called dideoxyribonucleotides (ddNTPs), are missing the 3´ hydroxyl group that is required for 5’ to 3’ extension of a DNA polynucleotide chain. By mixing ddNTPs that have been labeled with a different color for each base, unlabeled dNTPs, and template DNA in a polymerase-driven reaction, strands of each possible length are produced when the ddNTPs are randomly incorporated and terminate the chain. The extension products are then separated by electrophoresis, resolved to single-nucleotide differences in size. The chain-terminated fragments are detected by their fluorescent labels, with each color identifying one of the terminating ddNTPs. The sequence of the template DNA strand can thus be derived by analysis

3500 Genetic Analyzer is a capillary electrophoresis instrument. It has 8-capillary array and polymer DNA sequencing and/or fragment analysis reagents and consumables for system qualification. It has a Dell computer workstation with flat-screen monitor and integrated software for instrument control, data collection, quality control, and auto-analysis of sample files for base calling and fragment sizing