キャピラリー電気泳動によるDNAシークエンシング反応生成物の高速分離

概要

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The effect of the gel composition, electric field strength, and capillary length on the separation of DNA sequencing reaction product was investigated in order to achieve high-speed DNA sequencing on a large-scale for the Human Genome Project. In this study, we prepared gel-filled capillaries differing in gel concentration (3%T〜6%T), degree of crosslinking (0%C〜5%C), and capillary length (15.3cm〜36.3cm). Fluorescence-labeled A-termination sequencing reaction product (M13mp18 template) was first separated by capillary electrophoresis using the gel-filled capillaries prepared here in electric fields from 200V/cm to 414V/cm and then detected at 560nm by using an on-line argon ion laser (488nm)-induced fluorescence detector. In conclusion, the use of the short capillary, high concentration gel, and high electric field realizes high-speed separation of DNA sequencing reaction product with high resolution. Under optimum separation conditions, only 10 min is required for sequencing of 250-base DNA.
社団法人日本分析化学会の論文
1994-12-05