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.

Subject headings

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Keyword

DNA sequencing; primer-dimers; pyrosequencing technology; sample preparation; single-strand binding protein; SINGLE-NUCLEOTIDE POLYMORPHISM; TUMOR-SUPPRESSOR GENE; STRANDED DNA-BINDING; SEQUENCING PRIMERS; MUTATION; PROTEIN

Biochemistry

Biokemi

Publication and Content Type

ref (subject category)

art (subject category)