Quantification of Global DNA Methylation Levels by Mass Spectrometry

We describe the protocol that we currently use in our laboratory to analyze 5mC and 5hmC by mass spectrometry

Agustin F. Fernandez; Luis Valledor; Fernando Vallejo; Maria Jesús Cañal; Mario F. Fraga

2017

Scholarcy highlights

  • Global DNA methylation was classically considered the relative percentage of 5-methylcysine with respect to total cytosine
  • The recent development of protocols using mass spectrometry for the detection has increased sensibility and permitted the precise identification of peak compounds based on their molecular masses
  • We describe the protocol that we currently use in our laboratory to analyze 5mC and 5hmC by mass spectrometry
  • The protocol, which is based on the method originally developed by Le and colleagues using Ultra Performance Liquid Chromatography and mass spectrometry) detection, allows for the rapid and accurate quantification of relative global 5mC and 5hmC levels starting from just 1 μg of genomic DNA, which allows for the rapid and accurate quantification of relative global 5mC and 5hmC levels

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