Double-stranded DNA sequence analysis.

University of California Los Angeles, Los Angeles, California, USA.

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.

PMID: 18432690 [PubMed - indexed for MEDLINE]