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The Help Assay

  • Mayumi Oda
  • John M. Greally
Part of the Methods in Molecular Biology book series (MIMB, volume 507)

Abstract

Genomic representations using ligation-mediated PCR have been used successfully as the foundation for a number of high-throughput assays. HpaII tiny fragment enrichment by ligation-mediated PCR (HELP) is an example of the use of such representations to study cytosine methylation in the genome. The HELP assay differs from most other assays testing cytosine methylation because of its positive representation of hypomethylated DNA in the genome, whereas other assays infer the presence of hypomethylated sequences by the absence of signal, for which there can be confounding technical reasons. Hypomethylated sequences represent the minority of the genome and tend to be located at unique sequences with functionally interesting properties such as transcription start sites. By performing a comparative genomic hybridization using an MspI representation from the same DNA sample, we represent all potential loci that could be generated by HpaII in the situation of global hypomethylation; in practice, HpaII generates a subset of loci from this population, allowing us to discriminate hypomethylated loci (represented by both HpaII and MspI) from methylated loci (represented by MspI only).

Keywords

Cytosine methylation CG dinucleotide microarray epigenetic epigenome 

Notes

Acknowledgements

This work was originally supported by a grant from the National Institutes of Health (NCI) R03 CA111577 and subsequently by NICHD grant R01 HD044078 to JMG. The author also acknowledges the contributions of the Bioinformatics Shared Resource and the Genomics Core Facility at the Albert Einstein College of Medicine, as well as resources from the Albert Einstein Cancer Center.

References

  1. 1.
    Khulan, B., Thompson, R. F., Ye, K., et al. (2006) Comparative isoschizomer profiling of cytosine methylation: the HELP assay. Genome Res 16, 1046–1055.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Mayumi Oda
    • 1
  • John M. Greally
  1. 1.Departments of Medicine (Hematology) and Molecular GeneticsAlbert Einstein College of MedicineBronxUSA

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