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Comparative isoschizomer profiling of cytosine methylation: The HELP assay

Open access
Khulan, Batbayar, Thompson, Reid F., Ye, Kenny, Fazzari, Melissa J., Suzuki, Masako, Stasiek, Edyta, Figueroa, Maria E., Glass, Jacob L., Chen, Quan, Montagna, Cristina, Hatchwell, Eli, Selzer, Rebecca R., Richmond, Todd A., Green, Roland D., Melnick, Ari, Greally, John M.·2006·Genome Research

Abstract

The distribution of cytosine methylation in 6.2 Mb of the mouse genome was tested using cohybridization of genomic representations from a methylation-sensitive restriction enzyme and its methylation-insensitive isoschizomer. This assay, termed HELP (H paII tiny fragment E nrichment by L igation-mediated P CR), allows both intragenomic profiling and intergenomic comparisons of cytosine methylation. The intragenomic profile shows most of the genome to be contiguous methylated sequence with occasional clusters of hypomethylated loci, usually but not exclusively at promoters and CpG islands. Intergenomic comparison found marked differences in cytosine methylation between spermatogenic and brain cells, identifying 223 new candidate tissue-specific differentially methylated regions (T-DMRs). Bisulfite pyrosequencing confirmed the four candidates tested to be T-DMRs, while quantitative RT-PCR for two genes with T-DMRs located at their promoters showed the HELP data to be correlated with gene activity at these loci. The HELP assay is robust, quantitative, and accurate and is providing new insights into the distribution and dynamic nature of cytosine methylation in the genome.

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Metadata

Authors
Khulan, B., Thompson, R. F., Ye, K., Fazzari, M. J., Suzuki, M., Stasiek, E., Figueroa, M. E., Glass, J. L., Chen, Q., Montagna, C., Hatchwell, E., Selzer, R. R., Richmond, T. A., Green, R. D., Melnick, A., & Greally, J. M.
Venue
Genome Research
Published
2006-06-29
Volume
16
Issue
8
Pages
1046-1055
ISSN
1088-9051 (ISSN-L)
OpenAlex
W2112448007
Cited by
396
References
6
Type
posted content
Language
en
Open access
bronze

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CoChat. (2026). Citation check: Comparative isoschizomer profiling of cytosine methylation: The HELP assay. https://app.cochat.ai/papers/10.1101/gr.5273806
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Khulan, B., Thompson, R. F., Ye, K., Fazzari, M. J., Suzuki, M., Stasiek, E., Figueroa, M. E., Glass, J. L., Chen, Q., Montagna, C., Hatchwell, E., Selzer, R. R., Richmond, T. A., Green, R. D., Melnick, A., & Greally, J. M. (2006). Comparative isoschizomer profiling of cytosine methylation: The HELP assay. Genome Research. https://doi.org/10.1101/gr.5273806

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