High sensitivity mapping of methylated cytosines

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ID: 291193
1994
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Abstract
An understanding of DNA methylation and its potential role in gene control during development, aging and cancer has been hampered by a lack of sensitive methods which can resolve exact methylation patterns from only small quantities of DNA. We have now developed a genomic sequencing technique which is capable of detecting every methylated cytosine on both strands of any target sequence, using DNA isolated from fewer than 100 cells. In this method, sodium bisulphite is used to convert cytosine residues to uracil residues in single-stranded DNA, under conditions whereby 5-methylcytosine remains non-reactive. The converted DNA is amplified with specific primers and sequenced. All the cytosine residues remaining in the sequence represent previously methylated cytosines in the genome. The work described has defined procedures that maximise the efficiency of denaturetion, bisulphite conversion and amplification, to permit methylation mapping of single genes from small amounts of genomic DNA, readily available from germ cells and early developmental stages.
Reference Key
openalex_W1968263601 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors J.CIark Susan, Ewen M. Harrison, Cheryl Paul, Marianne Frommer
Journal Nucleic Acids Research
Year 1994
DOI
10.1093/nar/22.15.2990
URL
https://doi.org/10.1093/nar/22.15.2990
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