Methodological improvements of pyrosequencing technology

Baback Gharizadeh, Michael Akhras, Nader Nourizad, Mehran Ghaderi, Kenji Yasuda, Pål Nyrén, Nader Pourmand*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)504-511
Number of pages8
JournalJournal of Biotechnology
Issue number3
Publication statusPublished - 2006 Jul 25
Externally publishedYes


  • DNA sequencing
  • Primer-dimers
  • Pyrosequencing technology
  • Sample preparation
  • Single-strand binding protein

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


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