Separation of DNA sequencing fragments at 53 bases/minute by capillary gel electrophoresis

Abstract

Abstract Polyacrylamide gels are unstable in 50‐μm internal diameter capillaries at electric fields greater than about 500 V cm −1 due to the formation of bubbles, which destroy the separation. We describe the use of a modified polyacrylamide capillary gel that is stable at electric fields of at least 800 V cm −1 . This capillary gel produces sequencing rates of 3,200 bases h −1 for fragments up to 250 bases in length; a total of 7 min are required to separate fragments of 250 bases in length. Longer fragments tend to coelute and do not generate useful sequencing information. This sequencing rate is a factor of 40 times higher than that produced by conventional sequencing gels and a factor of 3 faster than previous reports in the literature.