Microarray-based DNA methylation profiling: Technology and applications

Axel Schumacher, Philipp Kapranov, Zachary A Kaminsky, James Flanagan, Abbas Assadzadeh, Patrick Yau, Carl Virtanen, Neil Winegarden, Jill Cheng, Thomas Gingeras, Arturas Petronis

Research output: Contribution to journalArticle

Abstract

This work is dedicated to the development of a technology for unbiased, high-throughput DNA methylation profiling of large genomic regions. In this method, unmethylated and methylated DNA fractions are enriched using a series of treatments with methylation sensitive restriction enzymes, and interrogated on microarrays. We have investigated various aspects of the technology including its replicability, informativeness, sensitivity and optimal PCR conditions using microarrays containing oligonucleotides representing 100 kb of genomic DNA derived from the chromosome 22 COM region in addition to 12 192 element CpG island microarrays. Several new aspects of methylation profiling are provided, including the parallel identification of confounding effects of DNA sequence variation, the description of the principles of microarray design for epigenomic studies and the optimal choice of methylation sensitive restriction enzymes. We also demonstrate the advantages of using the unmethylated DNA fraction versus the methylated one, which substantially improve the chances of detecting DNA methylation differences. We applied this methodology for fine-mapping of methylation patterns of chromosomes 21 and 22 in eight individuals using tiling microarrays consisting of over 340 000 oligonucleotide probe pairs. The principles developed in this work will help to make epigenetic profiling of the entire human genome a routine procedure.

Original languageEnglish (US)
Pages (from-to)528-542
Number of pages15
JournalNucleic Acids Research
Volume34
Issue number2
DOIs
StatePublished - Feb 2006
Externally publishedYes

Fingerprint

DNA Fingerprinting
DNA Methylation
Methylation
Technology
Chromosomes, Human, Pair 22
Epigenomics
DNA
Chromosomes, Human, Pair 21
CpG Islands
Oligonucleotide Probes
Human Genome
Enzymes
Oligonucleotide Array Sequence Analysis
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Genetics

Cite this

Schumacher, A., Kapranov, P., Kaminsky, Z. A., Flanagan, J., Assadzadeh, A., Yau, P., ... Petronis, A. (2006). Microarray-based DNA methylation profiling: Technology and applications. Nucleic Acids Research, 34(2), 528-542. https://doi.org/10.1093/nar/gkj461

Microarray-based DNA methylation profiling : Technology and applications. / Schumacher, Axel; Kapranov, Philipp; Kaminsky, Zachary A; Flanagan, James; Assadzadeh, Abbas; Yau, Patrick; Virtanen, Carl; Winegarden, Neil; Cheng, Jill; Gingeras, Thomas; Petronis, Arturas.

In: Nucleic Acids Research, Vol. 34, No. 2, 02.2006, p. 528-542.

Research output: Contribution to journalArticle

Schumacher, A, Kapranov, P, Kaminsky, ZA, Flanagan, J, Assadzadeh, A, Yau, P, Virtanen, C, Winegarden, N, Cheng, J, Gingeras, T & Petronis, A 2006, 'Microarray-based DNA methylation profiling: Technology and applications', Nucleic Acids Research, vol. 34, no. 2, pp. 528-542. https://doi.org/10.1093/nar/gkj461
Schumacher, Axel ; Kapranov, Philipp ; Kaminsky, Zachary A ; Flanagan, James ; Assadzadeh, Abbas ; Yau, Patrick ; Virtanen, Carl ; Winegarden, Neil ; Cheng, Jill ; Gingeras, Thomas ; Petronis, Arturas. / Microarray-based DNA methylation profiling : Technology and applications. In: Nucleic Acids Research. 2006 ; Vol. 34, No. 2. pp. 528-542.
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