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Methylation-Specific Multiplex Ligation-Dependent Probe Amplification (MS-MLPA).

Cathy B Moelans1, Lilit Atanesyan2, Suvi P Savola2

  • 1Department of Pathology, University Medical Centre Utrecht, Heidelberglaan 100, PO Box 85500, Utrecht, 3508 GA, The Netherlands. cmoelans@umcutrecht.nl.

Methods in Molecular Biology (Clifton, N.J.)
|December 11, 2017
PubMed
Summary
This summary is machine-generated.

This study presents methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) for rapid DNA methylation analysis. This PCR-based method efficiently detects promoter hypermethylation and imprinted region methylation without bisulfite conversion.

Keywords:
Coffalyser.NetFFPEGenomic DNAMLPAMS-MLPAMethylationMethylation-sensitive restriction enzymes paraffin

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Area of Science:

  • Molecular Biology
  • Genetics
  • Epigenetics

Background:

  • DNA methylation is crucial for gene regulation and implicated in diseases.
  • Accurate assessment of methylation patterns is vital for diagnostics and research.
  • Existing methods can be time-consuming or require large DNA amounts.

Purpose of the Study:

  • To describe a rapid and efficient method for assessing DNA methylation levels.
  • To introduce Methylation-Specific Multiplex Ligation-dependent Probe Amplification (MS-MLPA) as a diagnostic tool.
  • To enable the analysis of promoter hypermethylation and imprinted region methylation.

Main Methods:

  • Utilizes Multiplex Ligation-dependent Probe Amplification (MLPA), a PCR-based technique.
  • Employs the methylation-sensitive endonuclease HhaI, avoiding sodium bisulfite conversion.
  • Probes target CpG dinucleotides within CpG islands, with HhaI cutting unmethylated sites.

Main Results:

  • MS-MLPA allows rapid assessment of methylation in various human genomic DNA sources.
  • The method requires minimal DNA quantities (approx. 50 ng).
  • Methylation percentage is calculated by comparing HhaI-treated and untreated reactions.

Conclusions:

  • MS-MLPA provides a robust and efficient alternative for DNA methylation analysis.
  • The technique can be combined with copy number and point mutation detection.
  • MS-MLPA facilitates the study of epigenetic alterations in various biological samples.