Unit

UNIT 10.25 Double-Stranded DNA Sequence Analysis

  1. Sherie L. Morrison

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im1025s14

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Morrison, S. L. 2001. Double-Stranded DNA Sequence Analysis. Current Protocols in Immunology. 14:VII:10.25:10.25.1–10.25.7.

Author Information

  1. University of California Los Angeles, Los Angeles, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1995

Abstract

There are several different methods for analyzing the sequence of cloned DNA segments. For analysis of immunoglobulin variable (V) regions, double-stranded DNA sequencing is routinely used. Although this method does not yield the longest DNA sequences, it is easy and provides sufficient information to determine the correct sequence. The protocols in this unit are for use with the Sequenase Version 2.0 kit from U.S. Biochemicals, which is based on dideoxy sequencing with a modified T7 DNA polymerase. The Basic Protocol describes performing a sequencing reaction for cloned immunoglobulin V regions and running and analyzing a sequencing gel. A support protocol describes pouring and setting up the sequencing gel.