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High-resolution separation of oligonucleotides and DNA sequencing reaction products by capillary electrophoresis with linear polyacrylamide and laser-induced fluorescence detection

✍ Scribed by Nong Chen; Takashi Manabe; Shigeru Terabe; Masafumi Yohda; Isao Endo

Publisher
John Wiley and Sons
Year
1994
Tongue
English
Weight
426 KB
Volume
6
Category
Article
ISSN
1040-7685

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✦ Synopsis

Abstract

By employing linear polyacrylamide (LPA) as a sieving matrix, oligonucleotides and DNA sequencing fragments have been separated by capillary electrophoresis. A commercially available apparatus equipped with a laser‐induced fluorescence (LIF) detection system has been utilized, but the capillary cartridge has been modified in order to set the capillaries without coiling. Single‐base resolution was achieved for a mixture of FITC‐labeled oligonucleotides [FITC‐p(dT)] up to 500 base‐long chains. Because of high resolution, minor components of FITC‐p(dT) have also been separated. The mobility differences between single‐base adjacent peaks of FITC‐labeled DNA sequencing fragments have been detected up to 520 base‐long fragments. These results confirmed that linear polyacrylamide is an excellent sieving matrix in DNA sequencing by capillary electrophoresis.

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