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

✍ Scribed by Marie-Josée Rocheleau; Norman J. Dovichi

• Publisher: John Wiley and Sons
• Year: 1992
• Tongue: English
• Weight: 384 KB
• Volume: 4
• Category: Article
• ISSN: 1040-7685
• DOI: 10.1002/mcs.1220040513

✦ Synopsis

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.