Unit

UNIT 7.5 DNA Sequencing by the Chemical Method

  1. Richard L. Eckert

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0705s17

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Eckert, R. L. 2001. DNA Sequencing by the Chemical Method. Current Protocols in Molecular Biology. 17:7.5:7.5.1–7.5.11.

Author Information

  1. Case Western Reserve University, Cleveland, Ohio

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1992

This is not the most recent version of the article. View current version (1 OCT 2011)

Abstract

This procedure is written for a novice with no chemical sequencing experience. It is assumed, however, that the investigator has experience with DNA fragment isolation and DNA labeling with [32P]dNTPs. A DNA fragment labeled at only one end with 32P or 35S is divided into four aliquots and subjected to base-specific modification reactions. Piperidine is then added to catalyze strand scission at the modified bases. Finally, the four reactions are subjected to electrophoresis on adjacent lanes of a high-resolution denaturing polyacrylamide gel. The gel is then autoradiographed and the sequence is read from the film.