UNIT 10.25 Double-Stranded DNA Sequence Analysis
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: JUN 1995
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 describes pouring and setting up the sequencing gel.