Use of an electrochemically labeled nucleotide terminator for known point mutation analysis
- 13 August 2003
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 24 (16) , 2749-2757
- https://doi.org/10.1002/elps.200305554
Abstract
A novel single nucleotide polymorphism (SNP) assay utilizing an electrochemically tagged chain terminator is described. The system employs the single-base extension (SBE) technique coupled to capillary gel electrophoresis with end-column electrochemical detection. A redox-labeled chain terminator, ferrocene-acycloATP, is used in the SBE reaction. When the mutation site corresponds to the labeled chain terminator, the extension product is rendered electroactive. The reaction mixture is subsequently separated by capillary gel electrophoresis and the extension product detected at the separation anode with sinusoidal voltammetry. This work demonstrates the first known SNP assay utilizing redox-active chain terminators coupled to electrochemical detection. The methodology presented could lead to a fast, simple, and cost-effective SNP scoring system.Keywords
This publication has 42 references indexed in Scilit:
- Flexible Use of High-Density Oligonucleotide Arrays for Single-Nucleotide Polymorphism Discovery and ValidationGenome Research, 2001
- Synthesis of the First Ferrocene-Labeled Dideoxynucleotide and Its Use for 3‘-Redox End-Labeling of 5‘-Modified Single-Stranded OligonucleotidesBioconjugate Chemistry, 2001
- A semi-automated system for analysis and storage of SNPsHuman Mutation, 2001
- High-Performance Multiplex SNP Analysis of Three Hemochromatosis-Related Mutations With Capillary Array Electrophoresis MicroplatesGenome Research, 2001
- A map of human genome sequence variation containing 1.42 million single nucleotide polymorphismsNature, 2001
- Flow Cytometry-Based Minisequencing: A New Platform for High-Throughput Single-Nucleotide Polymorphism Scoring: Volume 66, Number 2 (2000), pages 135–143Genomics, 2000
- Single-Strand Conformation Polymorphism Analysis by Capillary and Microchip Electrophoresis: A Fast, Simple Method for Detection of Common Mutations in BRCA1 and BRCA2Genomics, 2000
- Applications of constant denaturant capillary electrophoresis / high-fidelity polymerase chain reaction to human genetic analysisElectrophoresis, 1999
- Large-Scale Identification, Mapping, and Genotyping of Single-Nucleotide Polymorphisms in the Human GenomeScience, 1998
- Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms.Proceedings of the National Academy of Sciences, 1989