UNIT 7.5 DNA Sequencing by the Chemical Method
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: JAN 1992
This is not the most recent version of the article. View current version (1 OCT 2011)
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.