A comparison of fluorescent SSCP and denaturing HPLC for high throughput mutation scanning

1 June 2000

journal article
research article
Published by Hindawi Limited in Human Mutation

Vol. 15 (6) , 556-564
https://doi.org/10.1002/1098-1004(200006)15:6<556::aid-humu7>3.0.co;2-c

Abstract

We examined 67 different mutations in 16 different amplicons in a comparison of mutation detection by fluorescent single strand conformation polymorphism (F-SSCP) and by denaturing HPLC (DHPLC). F-SSCP was used to analyze fluorescent amplicons with internal size standards and automated fragment analysis (GeneScan, PE Applied Biosystems, Foster City, CA). In DHPLC, unlabelled amplicons were analyzed by reverse phase HPLC with fragment detection by absorbance at 260nm. Both methods had high sensitivity (95-100%) and specificity (100%). Overall, F-SSCP with external temperature control was the more sensitive method, but DHPLC was particularly useful for the rapid analysis of novel fragments.

Keywords

This publication has 19 references indexed in Scilit:

High-throughput single-strand conformation polymorphism analysis by automated capillary electrophoresis: Robust multiplex analysis and pattern-based identification of allelic variants
Human Mutation, 1999
Blind Analysis of Denaturing High-Performance Liquid Chromatography as a Tool for Mutation Detection
Genomics, 1998
Denaturing HPLC-Identified Novel FBN1 Mutations, Polymorphisms, and Sequence Variants in Marfan Syndrome and Related Connective Tissue Disorders
Genetic Testing, 1997
Detection of Single-Nucleotide Polymorphisms with the WAVE™ DNA Fragment Analysis System
Genetic Testing, 1997
Comparison of Enzyme Mismatch Cleavage and Chemical Cleavage of Mismatch on a Defined Set of Heteroduplexes
Genetic Testing, 1997
Single-Strand Conformation Polymorphism (SSCP) Can Be Explained by Semistable Conformation Dynamics of Single-Stranded DNA
The Journal of Biochemistry, 1996
The Sensitivity of Single-Strand Conformation Polymorphism Analysis for the Detection of Single Base Substitutions
Genomics, 1993
Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms.
Proceedings of the National Academy of Sciences, 1989
Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations.
Proceedings of the National Academy of Sciences, 1988
[31] Detection and localization of single base changes by denaturing gradient gel electrophoresis
Published by Elsevier ,1987