Methylation specific oligonucleotide microarray
Methylation specific oligonucleotide microarray, also known as MSO microarray, was developed as a technique to map epigenetic methylation changes in DNA of cancer cells.[1]
The general process starts with modification of DNA with bisulfite, specifically to convert unmethylated cytosine in CpG sites to uracil, while leaving methylated cytosines untouched.[2] The modified DNA region of interest is amplified via PCR and during the process, uracils are converted to thymine. The amplicons are labelled with a fluorescent dye and hybridized to oligonucleotide probes that are fixed to a glass slide.[3] The probes differentially bind to cytosine and thymine residues, which ultimately allows discrimination between methylated and unmethylated CpG sites, respectively.[2]
A calibration curve is produced and compared with the microarray results of the amplified DNA samples. This allows a general quantification of the proportion of methylation present in the region of interest.[4]
This microarray technique was developed by Tim Hui-Ming Huang and his laboratory and was officially published in 2002.[5]
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Methylation specific oligonucleotide microarray was developed as a technique to map methylation changes in DNA in cancer. This technique was developed by Professor Tim Hui-Ming Huang and was published in journal Genome Research on 2002 (Gitan et al., 2002). The method utilizes bisulfite-modified DNA that is used as templates for PCR amplification, which is subsequently hybridized to oligonucleotide microarray.
External links
- Resources, information and specific protocols for DNA Methylation Analysis
- Software for DNA Methylation Analysis
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- ^ Gitan, Raad S.; Shi, Huidong; Chen, Chuan-Mu; Yan, Pearlly S.; Huang, Tim Hui-Ming (2002). "Methylation-Specific Oligonucleotide Microarray: A New Potential for High-Throughput Methylation Analysis". Genome Research. 12 (1): 158–164. doi:10.1101/gr.202801. ISSN 1088-9051. PMID 11779841.
- ^ a b Gitan, Raad S.; Shi, Huidong; Chen, Chuan-Mu; Yan, Pearlly S.; Huang, Tim Hui-Ming (2002). "Methylation-Specific Oligonucleotide Microarray: A New Potential for High-Throughput Methylation Analysis". Genome Research. 12 (1): 158–164. doi:10.1101/gr.202801. ISSN 1088-9051. PMID 11779841.
- ^ Shi, Huidong; Maier, Sabine; Nimmrich, Inko; Yan, Pearlly S.; Caldwell, Charles W.; Olek, Alexander; Huang, Tim Hui-Ming (2003-01-01). "Oligonucleotide-based microarray for DNA methylation analysis: principles and applications". Journal of Cellular Biochemistry. 88 (1): 138–143. doi:10.1002/jcb.10313. ISSN 0730-2312. PMID 12461783.
- ^ Yan, Pearlly S.; Wei, Susan H.; Huang, Tim Hui-Ming (2004), Tollefsbol, Trygve O. (ed.), "Methylation-Specific Oligonucleotide Microarray", Epigenetics Protocols, Methods in Molecular Biology™, Humana Press, pp. 251–260, doi:10.1385/1-59259-828-5:251, ISBN 9781592598281, retrieved 2019-10-05
- ^ Gitan, Raad S.; Shi, Huidong; Chen, Chuan-Mu; Yan, Pearlly S.; Huang, Tim Hui-Ming (2002). "Methylation-Specific Oligonucleotide Microarray: A New Potential for High-Throughput Methylation Analysis". Genome Research. 12 (1): 158–164. doi:10.1101/gr.202801. ISSN 1088-9051. PMID 11779841.