TY  - JOUR
A1  - Inazuka, Masakazu
A1  - Wenz, Hans-Michael
A1  - Sakabe, Munechika
A1  - Tahira, Tomoko
A1  - Hayashi, Kenshi
T1  - A Streamlined Mutation Detection System: Multicolor Post-PCR Fluorescence Labeling and Single-Strand Conformational Polymorphism Analysis by Capillary Electrophoresis
Y1  - 1997/11/01 
JF  - Genome Research 
JO  - Genome Research 
SP  - 1094 
EP  - 1103 
DO  - 10.1101/gr.7.11.1094 
VL  - 7 
IS  - 11 
UR  - http://genome.cshlp.org/content/7/11/1094.abstract 
N2  - Effective use of knowledge of human genome sequences in studies of hereditary diseases or cancer heavily depends on efficient methods for detection of mutations in individual samples. We describe here a simple and efficient mutation scanning system in which PCR products arepost-labeled with two different fluorescent dyes in one tube, and analyzed by an automatedcapillary electrophoresis system using single-strand conformation polymorphism (SSCP) conditions (PLACE–SSCP). With the appropriate use of an internal control DNA, differences in electrophoretic mobilities between a reference and samples are precisely evaluated, then the presence of mutations is statistically judged. Thirty-three of 34 known mutations in fragments of three unrelated sequence contexts up to 741 bp were detected using one electrophoresis condition at the confidence level of <0.3% false positive. All the mutations were detected by analyzing at two temperatures. The described system has the advantage of little human intervention, short analysis time, high sensitivity, and objectivity of data interpretation. 
ER  -