DNA Methylation Analysis Using Bisulfite Pyrosequencing.
Biomarker detection
Bisulfite conversion
CpG site
DNA methylation
Primer design
Pyrosequencing technology
SNPs
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2023
2023
Historique:
entrez:
29
9
2022
pubmed:
30
9
2022
medline:
4
10
2022
Statut:
ppublish
Résumé
Pyrosequencing is a DNA sequencing-by-synthesis technique that can quantitatively detect single-nucleotide polymorphisms (SNPs). With pyrosequencing, the level of DNA methylation can be calculated according to the ratio of artificial cytosine/thymine SNPs produced by bisulfite conversion at each CpG site. This analysis method provides a reproducible and accurate measurement of methylation levels at CpG sites near sequencing primers with high quantitative resolution. DNA methylation plays an important role in mammalian development and cellular physiology; alterations in DNA methylation patterns have been implicated in several common diseases as well as cancers and imprinting disorders. Evaluating DNA methylation levels via pyrosequencing is useful for identifying biomarkers that could help with the diagnosis, prognosis, treatment selection, and onset risk assessment for several diseases. We describe the principles of pyrosequencing and detail a bisulfite pyrosequencing protocol based on our experience and the PyroMark Q24 User Manual.
Identifiants
pubmed: 36173562
doi: 10.1007/978-1-0716-2724-2_1
doi:
Substances chimiques
DNA Primers
0
Sulfites
0
Cytosine
8J337D1HZY
hydrogen sulfite
OJ9787WBLU
Thymine
QR26YLT7LT
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3-20Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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