Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4118

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Single-Strand Conformation Polymorphism Analysis

Anne E. Jedlicka and Steven R. Kleeberger

This protocol was adapted from "Single-Strand Conformation Polymorphism Analysis," contributed by Anne E. Jedlicka and Steven R. Kleeberger, Chapter 18, in PCR Primer, 2nd edition (eds. Dieffenbach and Dveksler). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 100 words of the full text of this article appear below.


INTRODUCTION

Single-strand confirmation polymorphism analysis (SSCP) is a powerful and robust method for the detection of DNA sequence changes (single-base substitutions) based on shifts in electrophoretic mobility. In this protocol, the target sequence is simultaneously labeled and amplified, then heat-denatured and resolved by non-denaturing polyacrylamide gel electrophoresis. Differences in sequence alter the conformation of the DNA and hence its electrophoretic mobility and, because of the high resolution of polyacrylamide gels, most conformational changes caused by subtle changes in sequence can be detected (Hayashi 1991).


MATERIALS

Reagents

caution[{gamma}-32P]ATP, 6000 Ci/mM (Perkin Elmer Life Sciences, or equivalent)

recipe caution Ammonium persulfate 10%

recipe dNTP solution, 10 . . . [Full Text of this Article]

Equipment


METHOD


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