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Automatic matrix determination in four dye fluorescence‐based DNA sequencing
Author(s) -
Yin Zhongbin,
Severin Jessica,
Giddings Michael C.,
Huang Weian,
Westphall Michael S.,
Smith Lloyd M.
Publication year - 1996
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/elps.1150170626
Subject(s) - fluorescence , transformation (genetics) , matrix (chemical analysis) , multispectral image , biological system , computer science , transformation matrix , sequence (biology) , complex matrix , calibration , algorithm , chemistry , artificial intelligence , optics , physics , chromatography , biology , biochemistry , kinematics , classical mechanics , quantum mechanics , gene
The four dye fluorescence detection strategy is a widely used approach to automated DNA sequence analysis. An important aspect of data processing in this approach is the multicomponent analysis to deduce the concentrations of four fluorophores from fluorescence emission intensities at four different wave‐lengths. This requires knowledge of the correct transformation matrix M. The matrix M is a function both of the fluorophores employed and the fluorescence detection system. M is typically determined either by a calibration process with individual dyes, or by choosing four well‐separated individual peaks corresponding to the four different dyes. Both are time‐consuming and complicated procedures for routine use. An automatic scheme for finding M directly from raw sequence data is presented here. This facilitates data analysis and the underlying algorithm may also find utility in other multispectral applications.