ArticleBMC molecular biology2017
Methylated site display (MSD)-AFLP, a sensitive and affordable method for analysis of CpG methylation profiles.
Article in BMC molecular biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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7 citing papers in PubMed.
- Review
- An aberrant DNA methylation signature for predicting the prognosis of head and neck squamous cell carcinoma.Cancer medicine · 2021Article
- Application of 5-Methylcytosine DNA Glycosylase to the Quantitative Analysis of DNA Methylation.International journal of molecular sciences · 2021Article
- Exploring disease-specific methylated CpGs in human male genital abnormalities by using methylated-site display-amplified fragment length polymorphism (MSD-AFLP).The Journal of reproduction and development · 2019Article
- Linking inter-individual variability to endocrine disruptors: insights for epigenetic inheritance.Mammalian genome : official journal of the International Mammalian Genome Society · 2018Article
- Does the prenatal bisphenol A exposure alter DNA methylation levels in the mouse hippocampus?: An analysis using a high-sensitivity methylome technique.Genes and environment : the official journal of the Japanese Environmental Mutagen Society · 2018Article
- Detection of dioxin-induced demethylation of mouseGenes and environment : the official journal of the Japanese Environmental Mutagen Society · 2018Article
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Authors and funding
10 authors.
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Abstract
backgroundIt has been pointed out that environmental factors or chemicals can cause diseases that are developmental in origin. To detect abnormal epigenetic alterations in DNA methylation, convenient and cost-effective methods are required for such research, in which multiple samples are processed simultaneously. We here present methylated site display (MSD), a unique technique for the preparation of DNA libraries. By combining it with amplified fragment length polymorphism (AFLP) analysis, we developed a new method, MSD-AFLP.
resultsMethylated site display libraries consist of only DNAs derived from DNA fragments that are CpG methylated at the 5' end in the original genomic DNA sample. To test the effectiveness of this method, CpG methylation levels in liver, kidney, and hippocampal tissues of mice were compared to examine if MSD-AFLP can detect subtle differences in the levels of tissue-specific differentially methylated CpGs. As a result, many CpG sites suspected to be tissue-specific differentially methylated were detected. Nucleotide sequences adjacent to these methyl-CpG sites were identified and we determined the methylation level by methylation-sensitive restriction endonuclease (MSRE)-PCR analysis to confirm the accuracy of AFLP analysis. The differences of the methylation level among tissues were almost identical among these methods. By MSD-AFLP analysis, we detected many CpGs showing less than 5% statistically significant tissue-specific difference and less than 10% degree of variability. Additionally, MSD-AFLP analysis could be used to identify CpG methylation sites in other organisms including humans.
conclusionMSD-AFLP analysis can potentially be used to measure slight changes in CpG methylation level. Regarding the remarkable precision, sensitivity, and throughput of MSD-AFLP analysis studies, this method will be advantageous in a variety of epigenetics-based research.
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