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Improvements in Pyrosequencing technology by employing Sequenase polymerase
Stanford Genome Technology Center, Stanford University, Palo Alto, USA.
Department of Biotechnology, Stockholm Center for Physics, Astronomy and Biotechnology, Royal Institute of Technology, Stockholm.
Department of Biotechnology, Stockholm Center for Physics, Astronomy and Biotechnology, Royal Institute of Technology, Stockholm.
Department of Biotechnology, Stockholm Center for Physics, Astronomy and Biotechnology, Royal Institute of Technology, Stockholm.
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2004 (engelsk)Inngår i: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 330, nr 2, s. 272-280Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Pyrosequencing is a DNA sequencing technique based on the bioluminometric detection of inorganic pyrophosphate, which is released when nucleotides are incorporated into a target DNA. Since the technique is based on an enzymatic cascade, the choice of enzymes is a critical factor for efficient performance of the sequencing reaction. In this study we have analyzed the performance of an alternative DNA polymerase, Sequenase, on the sequencing performance of the Pyrosequencing technology. Compared to the Klenow fragment of DNA polymerase I, Sequenase could read through homopolymeric regions with more than five T bases. In addition, Sequenase reduces remarkably interference from primer-dimers and loop structures that give rise to false sequence signals. By using Sequenase, synchronized extensions and longer reads can be obtained on challenging templates, thereby opening new avenues for applications of Pyrosequencing technology.

sted, utgiver, år, opplag, sider
Academic Press, 2004. Vol. 330, nr 2, s. 272-280
Emneord [en]
pyrosequencing technology, sequenase, primer-dimers, loop structures, DNA sequencing
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Identifikatorer
URN: urn:nbn:se:umu:diva-85453DOI: 10.1016/j.ab.2004.03.018ISI: 000222424800012PubMedID: 15203333OAI: oai:DiVA.org:umu-85453DiVA, id: diva2:693578
Tilgjengelig fra: 2014-02-04 Laget: 2014-02-04 Sist oppdatert: 2018-06-08bibliografisk kontrollert

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