Methodological improvements of pyrosequencing technology.
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.
Gharizadeh
Baback
B
Stanford Genome Technology Center, Stanford University, 855 California Avenue, Palo Alto, CA 94304, USA.
Akhras
Michael
M
Nourizad
Nader
N
Ghaderi
Mehran
M
Yasuda
Kenji
K
Nyrén
Pål
P
Pourmand
Nader
N
D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*; D013048:Specimen Handling / Q000379:methods*