Direct analysis of single‐nucleotide polymorphism on double‐stranded DNA by pyrosequencing
- 1 April 2000
- journal article
- Published by Wiley in Biotechnology and Applied Biochemistry
- Vol. 31 (2), 107-112
- https://doi.org/10.1042/ba19990104
Abstract
Pyrosequencing, a new method for DNA sequencing, is gaining widespread use for many different types of DNA analysis. The method takes advantage of four coupled enzymes in a single tube assay to monitor DNA synthesis in real time using a luminometric detection system. Here, we demonstrate the use of pyrosequencing for direct analysis of single‐nucleotide polymorphism on double‐stranded PCR product. Pyrosequencing data on the human glutathione peroxidase gene (GPX1) from several individuals were analysed and three different allelic variants were determined and confirmed. The possibility of further simplifying the sequencing and template‐preparation steps is discussed.Keywords
This publication has 8 references indexed in Scilit:
- Production, Purification, and Luminometric Analysis of RecombinantSaccharomyces cerevisiae MET3Adenosine Triphosphate Sulfurylase Expressed inEscherichia coliProtein Expression and Purification, 1999
- Low yield of polymorphisms from EST blast searching: Analysis of genes related to oxidative stress and verification of the P197L polymorphism in GPX1Human Mutation, 1999
- Analyses of Secondary Structures in DNA by PyrosequencingAnalytical Biochemistry, 1999
- Reading Bits of Genetic Information: Methods for Single-Nucleotide Polymorphism AnalysisGenome Research, 1998
- A Sequencing Method Based on Real-Time PyrophosphateScience, 1998
- Large-Scale Identification, Mapping, and Genotyping of Single-Nucleotide Polymorphisms in the Human GenomeScience, 1998
- Increasing the Information Content of STS-Based Genome Maps: Identifying Polymorphisms in Mapped STSsGenomics, 1996
- An estimate of unique DNA sequence heterozygosity in the human genomeHuman Genetics, 1985