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DNA sequencing by capillary electrophoresis
Author(s) -
Dovichi Norman J.
Publication year - 1997
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.1150181229
Subject(s) - capillary electrophoresis , dna sequencing , dna , electrophoresis , gel electrophoresis , capillary action , genomic dna , electric field , biology , materials science , genetics , microbiology and biotechnology , physics , composite material , quantum mechanics
Capillary electrophoresis has been under development for DNA sequencing since 1990. This development has traveled down two parallel tracks. The first track studied the details of DNA separation by gel electrophoresis. Early work stressed rapid separations at high electric fields, which reached the extreme of a 3.5 min sequencing run at 1200 V/cm. While fast separations are useful in clinical resequencing applications for mutation detection, long read‐length is important in genomic sequencing. Unfortunately, sequence read‐length degrades as electric field and sequencing speed increases; this tradeoff between read‐length and sequencing speed appears to be a fundamental result of the physics of DNA separations in a polymer. The longest sequence sequencing read‐lengths have been obtained at modest electric fields, high temperature, and with low concentration noncrosslinked polymers. In parallel with our understanding of DNA separations, the second track of DNA sequencing development considered the design of large‐scale capillary instruments, wherein hundreds of DNA samples can be sequenced in parallel. Real‐world application of these very high throughput capillary electrophoresis systems will require significant investment in sample preparation technology.