Direct analysis of single-nucleotide polymorphism on double-stranded DNA by pyrosequencing

被引：55

作者：

Nordström, T

Ronaghi, M

Forsberg, L

de Faire, U

Morgenstern, R

Nyrén, P ^{[1
]}

机构：

[1] Royal Inst Technol, Dept Biotechnol, SE-10044 Stockholm, Sweden

[2] Stanford Univ, DNA Sequencing & Technol Ctr, Palo Alto, CA 94304 USA

[3] Karolinska Inst, Inst Environm Med, Div Biochem Toxicol, SE-17177 Stockholm, Sweden

[4] Karolinska Inst, Inst Environm Med, Div Cardiovasc Epidemiol, SE-17177 Stockholm, Sweden

来源：

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY | 2000年 / 31卷

关键词：

alkaline phosphatase; apyrase; exonuclease I; human glutathione peroxidase; inorganic pyrophosphate;

D O I：

10.1042/BA19990104

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (GPXI) 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.

引用

页码：107 / 112

页数：6

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