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Methylation-specific PCR.

Publication ,  Journal Article
Huang, Z; Bassil, CF; Murphy, SK
Published in: Methods Mol Biol
2013

Defining DNA methylation patterns in the genome has become essential for understanding diverse biological processes including the regulation of gene expression, imprinted genes, and X chromosome inactivation and how these patterns are deregulated in human diseases. Methylation-specific (MS)-PCR is a useful tool for qualitative DNA methylation analysis with multiple advantages, including ease of design and execution, sensitivity in the ability to detect small quantities of methylated DNA, and the ability to rapidly screen a large number of samples without the need for purchase of expensive laboratory equipment. This assay requires modification of the genomic DNA by sodium bisulfite and two independent primer sets for PCR amplification, one pair designed to recognize the methylated and the other pair the unmethylated versions of the bisulfite-modified sequence. The amplicons are visualized using ethidium bromide staining following agarose gel electrophoresis. Amplicons of the expected size produced from either primer pair are indicative of the presence of DNA in the original sample with the respective methylation status.

Duke Scholars

Published In

Methods Mol Biol

DOI

EISSN

1940-6029

Publication Date

2013

Volume

1049

Start / End Page

75 / 82

Location

United States

Related Subject Headings

  • Sulfites
  • Polymerase Chain Reaction
  • Humans
  • Genome, Human
  • Ethidium
  • Developmental Biology
  • DNA Primers
  • DNA Methylation
  • DNA
  • CpG Islands
 

Citation

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ICMJE
MLA
NLM
Huang, Z., Bassil, C. F., & Murphy, S. K. (2013). Methylation-specific PCR. Methods Mol Biol, 1049, 75–82. https://doi.org/10.1007/978-1-62703-547-7_7
Huang, Zhiqing, Christopher F. Bassil, and Susan K. Murphy. “Methylation-specific PCR.Methods Mol Biol 1049 (2013): 75–82. https://doi.org/10.1007/978-1-62703-547-7_7.
Huang Z, Bassil CF, Murphy SK. Methylation-specific PCR. Methods Mol Biol. 2013;1049:75–82.
Huang, Zhiqing, et al. “Methylation-specific PCR.Methods Mol Biol, vol. 1049, 2013, pp. 75–82. Pubmed, doi:10.1007/978-1-62703-547-7_7.
Huang Z, Bassil CF, Murphy SK. Methylation-specific PCR. Methods Mol Biol. 2013;1049:75–82.

Published In

Methods Mol Biol

DOI

EISSN

1940-6029

Publication Date

2013

Volume

1049

Start / End Page

75 / 82

Location

United States

Related Subject Headings

  • Sulfites
  • Polymerase Chain Reaction
  • Humans
  • Genome, Human
  • Ethidium
  • Developmental Biology
  • DNA Primers
  • DNA Methylation
  • DNA
  • CpG Islands