Automated high-accuracy mutation screening with the WAVE nucleic acid fragment analysis system

Karl Heinz Hecker

doi:10.1117/12.472095

21 June 2002 Automated high-accuracy mutation screening with the WAVE nucleic acid fragment analysis system

Karl Heinz Hecker

Proceedings Volume 4626, Biomedical Nanotechnology Architectures and Applications; (2002) https://doi.org/10.1117/12.472095
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

The analysis of DNA fragments by ion-pair reversed-phase high-performance liquid chromatography on an alkylated, nonporous poly(styrene-divinylbenzene) matrix (DNA Cartridge) using the WAVE Nucleic Acid Fragment Analysis System is a powerful and versatile tool for DNA analysis. Resolution of DNA fragments is based on two principles, size-dependent retention of double-stranded (ds) DNA and differential retention of ds vs. single-stranded (ss) DNA. Temperature Modulated Heteroduplex Analysis utilizes both principles of separation to detect single nucleotide polymorphisms (SNP) and short insertions/deletions. At a given temperature the difference in the melting between homo- and heteroduplexes is revealed by differences in retention times. The temperature at which differential melting occurs is sequence dependent and is predicated accurately using either WAVEMAKER or WAVE Navigator software, which use a modified Fixman-Friere algorithm. Detection of known and unknown sequence variations can be performed on DNA fragments of up to 1,000 base pairs with high sensitivity and specificity. The use of fluorescent labels is compatible with the technology and increases sensitivity. Retention times are increased and resolution is not affected. Fluorescent labeling significantly increases sensitivity.

Citation Download Citation

Karl Heinz Hecker "Automated high-accuracy mutation screening with the WAVE nucleic acid fragment analysis system", Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); https://doi.org/10.1117/12.472095

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