Pyrosequencing analysis for breast cancer DNA methylome

Cem Kuscu, Canan Kuscu

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Unraveling DNA methylation profile of tumor is important for the diagnosis and treatment of cancer patients. Because of the heterogeneity of clinical samples, it is very difficult to get methylation profile of only tumor cells. Laser capture Microdissection (LCM) is giving us a chance to isolate the DNA only from the tumor cells without any stroma cell’s DNA contamination. Once we capture the breast tumor cells, we can isolate the genomic DNA which is followed by the bisulfite treatment in which unmethylated cytosines of the CG pairs are converted into uracil; however, methylated cytosine does not go into any chemical change during this reaction. Next, bisulfite treated DNA is used in the regular PCR reaction to get a single band PCR amplicon which will be used as a template for the pyrosequencing. Pyrosequencing is a powerful method to make a quantitative methylation analysis for each specific CG pair.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages89-104
Number of pages16
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1406
ISSN (Print)1064-3745

Fingerprint

Breast Neoplasms
Cytosine
DNA
Methylation
Neoplasms
DNA Contamination
Laser Capture Microdissection
Polymerase Chain Reaction
Uracil
DNA Methylation
Therapeutics
hydrogen sulfite

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Kuscu, C., & Kuscu, C. (2016). Pyrosequencing analysis for breast cancer DNA methylome. In Methods in Molecular Biology (pp. 89-104). (Methods in Molecular Biology; Vol. 1406). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3444-7_8

Pyrosequencing analysis for breast cancer DNA methylome. / Kuscu, Cem; Kuscu, Canan.

Methods in Molecular Biology. Humana Press Inc., 2016. p. 89-104 (Methods in Molecular Biology; Vol. 1406).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kuscu, C & Kuscu, C 2016, Pyrosequencing analysis for breast cancer DNA methylome. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1406, Humana Press Inc., pp. 89-104. https://doi.org/10.1007/978-1-4939-3444-7_8
Kuscu C, Kuscu C. Pyrosequencing analysis for breast cancer DNA methylome. In Methods in Molecular Biology. Humana Press Inc. 2016. p. 89-104. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-3444-7_8
Kuscu, Cem ; Kuscu, Canan. / Pyrosequencing analysis for breast cancer DNA methylome. Methods in Molecular Biology. Humana Press Inc., 2016. pp. 89-104 (Methods in Molecular Biology).
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