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Plant Epigenetics

Volume 631 of the series Methods in Molecular Biology™ pp 1-11

Date:

Analysis of DNA Methylation in Plants by Bisulfite Sequencing

  • Andrea M. FoersterAffiliated withGregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences
  • , Ortrun Mittelsten ScheidAffiliated withGregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences Email author 

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Abstract

Methylation of cytosines is a very important epigenetic modification of genomic DNA in many different eukaryotes, and it is frequently involved in transcriptional regulation of genes. In plants, DNA methylation is regulated by a complex interplay between several methylating and demethylating enzymes. Analysis of the resulting cytosine methylation patterns with the highest resolution is achieved after sodium bisulfite treatment, deaminating nonmethylated cytosines to uracil. Subsequent PCR and sequence analysis of individual amplicons displays the degree, position, and sequence context of methylation of every cytosine residue in individual genomic sequences. We describe the application of bisulfite sequencing for the analysis of DNA methylation at defined individual sequences of plant genomic DNA.

Key words

DNA methylation 5-methylcytosine (5mC) Bisulfite sequencing Bisulfite primer design Bisulfite conversion control