Appendix

APPENDIX 3B Denaturing Polyacrylamide Gel Electrophoresis

  1. Lisa M. Albright1,
  2. Barton E. Slatko2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142700.nca03bs00

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Albright, L. M. and Slatko, B. E. 2001. Denaturing Polyacrylamide Gel Electrophoresis. Current Protocols in Nucleic Acid Chemistry. 00:3B:A.3B.1–A.3B.5.

Author Information

  1. 1

    Allison Park, Pennsylvania

  2. 2

    New England Biolabs, Beverly, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: FEB 2000

Abstract

Thin polyacrylamide gels that contain a high concentration of urea as a denaturant are capable of resolving short (<500 nucleotides) single-stranded fragments of DNA or RNA that differ in length by as little as one nucleotide. Such gels are uniquely suited for nucleic acid sequence analysis, which is required, for instance, for all footprinting protocols. Thicker gels are often used to purify oligonucleotides. This appendix describes the pouring, running, and processing of a typical sequencing gel, which is 40 cm long with a uniform thickness of 0.4 mm, containing 7 M urea and 4% to 8% acrylamide.