Dynamic range of fluorescence detection and base-calling accuracy in DNA sequencer based on single-photon counting

• Published in: Electrophoresis Vol. 24; no. 7-8; pp. 1184 - 1192
• Main Authors: Gavrilov, Dmitri N., Gorbovitski, Boris, Gouzman, Mikhail, Gudkov, Georgiy, Stepoukhovitch, Andrei, Ruskovoloshin, Victor, Tsuprik, Andrei, Tyshko, Georgiy, Bilenko, Olga, Kosobokova, Olga, Luryi, Serge, Gorfinkel, Vera
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.04.2003; WILEY‐VCH Verlag
• Subjects: DNA sequencing instruments; Electronics; Electrophoresis, Capillary - instrumentation; Electrophoresis, Capillary - standards; Equipment Design; Fluorescent Dyes - analysis; Nucleotides - analysis; Photons; Reference Values; Reproducibility of Results; Sequence Analysis, DNA - instrumentation; Sequence Analysis, DNA - standards; Single-photon detection
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.200390152

Recently, we developed a family of high‐performance automated capillary DNA sequencing instruments based on a single‐photon detection of fluorescently labeled DNA fragments. Our machines employ digital and broadband techniques, essential for achieving superior instrument sensitivity and dynamic range. In the present paper, we discuss limitations of the instrument's performance caused by the nonlinearity of single‐photon detectors as well as methods for nonlinearity compensation which increase the detection dynamic range and base‐calling accuracy.