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Direct analysis of single-nucleotide polymorphism on double-stranded DNA by pyrosequencing
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2000 (English)In: Biotechnology and applied biochemistry, ISSN 0885-4513, E-ISSN 1470-8744, Vol. 31, 107-112 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2000. Vol. 31, 107-112 p.
Keyword [en]
alkaline phosphatase, apyrase, exonuclease I, human glutathione peroxidase, inorganic pyrophosphate, human genome, information
URN: urn:nbn:se:kth:diva-19705ISI: 000086605300006OAI: diva2:338397
QC 20100525Available from: 2010-08-10 Created: 2010-08-10Bibliographically approved

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