TY - JOUR
T1 - Methodological improvements of pyrosequencing technology
AU - Gharizadeh, Baback
AU - Akhras, Michael
AU - Nourizad, Nader
AU - Ghaderi, Mehran
AU - Yasuda, Kenji
AU - Nyrén, Pål
AU - Pourmand, Nader
N1 - Funding Information:
We would like to thank Claus C. Hübner for his excellent technical assistance and Julianna P. Erickson for critical reading of this work. This study was supported, in part, by a grant from the National Institutes of Health, 1R21 A1059499-01 and PO1-HG000205.
PY - 2006/7/25
Y1 - 2006/7/25
N2 - Pyrosequencing technology is a rather novel DNA sequencing method based on the sequencing-by-synthesis principle. This bioluminometric, real-time DNA sequencing technique employs a cascade of four enzymatic reactions producing sequence peak signals. The method has been proven highly suitable for single nucleotide polymorphism analysis and sequencing of short stretches of DNA. Although the pyrosequencing procedure is relatively straightforward, users may face challenges due to varying parameters in PCR and sequencing primer design, sample preparation and nucleotide dispensation; such challenges are labor and cost intensive. In this study, these issues have been addressed to increase signal quality and assure sequence accuracy.
AB - Pyrosequencing technology is a rather novel DNA sequencing method based on the sequencing-by-synthesis principle. This bioluminometric, real-time DNA sequencing technique employs a cascade of four enzymatic reactions producing sequence peak signals. The method has been proven highly suitable for single nucleotide polymorphism analysis and sequencing of short stretches of DNA. Although the pyrosequencing procedure is relatively straightforward, users may face challenges due to varying parameters in PCR and sequencing primer design, sample preparation and nucleotide dispensation; such challenges are labor and cost intensive. In this study, these issues have been addressed to increase signal quality and assure sequence accuracy.
KW - DNA sequencing
KW - Primer-dimers
KW - Pyrosequencing technology
KW - Sample preparation
KW - Single-strand binding protein
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U2 - 10.1016/j.jbiotec.2006.01.025
DO - 10.1016/j.jbiotec.2006.01.025
M3 - Article
C2 - 16530870
AN - SCOPUS:33745504382
SN - 0168-1656
VL - 124
SP - 504
EP - 511
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 3
ER -