A tag-based approach for high-throughput analysis of CCWGG methylation |
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Authors: | Denisova Oksana V Chernov Andrei V Koledachkina Tatyana Y Matvienko Nicholas I |
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Institution: | Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Russian Federation. |
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Abstract: | Non-CpG methylation occurring in the context of CNG sequences is found in plants at a large number of genomic loci. However, there is still little information available about non-CpG methylation in mammals. Efficient methods that would allow detection of scarcely localized methylated sites in small quantities of DNA are required to elucidate the biological role of non-CpG methylation in both plants and animals. In this study, we tested a new whole genome approach to identify sites of CCWGG methylation (W is A or T), a particular case of CNG methylation, in genomic DNA. This technique is based on digestion of DNAs with methylation-sensitive restriction endonucleases EcoRII-C and AjnI. Short DNAs flanking methylated CCWGG sites (tags) are selectively purified and assembled in tandem arrays of up to nine tags. This allows high-throughput sequencing of tags, identification of flanking regions, and their exact positions in the genome. In this study, we tested specificity and efficiency of the approach. |
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Keywords: | Non-CpG methylation DNA methylation CCWGG methylation Epigenome |
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