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Alternative base‐calling algorithm for DNA sequencing based on four‐label multicolor detection
Author(s) -
Song Joon Myong,
Yeung Edward S.
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000301)21:4<807::aid-elps807>3.0.co;2-y
Subject(s) - dna , dna sequencing , base (topology) , capillary electrophoresis , resolution (logic) , high resolution , detection limit , base pair , chromatography , computer science , analytical chemistry (journal) , chemistry , artificial intelligence , biology , genetics , mathematics , remote sensing , mathematical analysis , geology
A simple base‐calling scheme based on four‐label multicolor detection is suggested for DNA sequencing. The entire spectra of the dye labels were used for identification. Specifically, the maxima of the emission spectra rather than the intensity ratios at selected wavelengths are used to provide excellent discrimination. Capillary gel electrophoresis was used for the separation of DNA fragments. Data acquisition and analysis compatible with fast and high‐throughput imaging detection was accomplished. The accuracy of base calling of PGEM/U DNA from the raw data obtained with 5 nm and 7 nm spectroscopic resolution were 98.4% for 386 bases and 98.4% for 385 bases. Base calling of M13mp18 DNA showed 98.3% accuracy for 420 bases.