Post-electrophoresis capillary scanner for detecting DNA sequencing reaction products

Abstract

A post-electrophoresis capillary scanning technique has been developed and its feasibility as a high-throughput DNA sequencing method has been demonstrated. In this method, capillary gel electrophoresis separations of DNA sequencing reaction products are spatially and temporally separated with laser-induced fluorescence detection of the products. A 90-cm gel-filled capillary was used to separate a mixture of the fluorescently labeled adenine-terminated DNA fragments of pBluescript SK(-) DNA. The adenine base sequence of the DNA was then determined up to 350 bases with a scan rate of 3.7 cm/s, which took 27 s for the capillary. We have estimated that the capillary scanner can analyze over 130 capillaries in 1 h, which corresponds to a DNA sequencing throughput of about 1 megabase/day.