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A Modified and Improved Method for Bisulphite Based Cytosine Methylation Analysis
Open accessAbstract
Sequencing of bisulphite modified genomic DNA is the most powerful method to determine methylation patterns in chromosomal DNA. In many experimental systems, the amount of material available for analysis is very small which makes it necessary to perform experiments at extreme levels of sensitivity and reproducibility. In this communication, we present an improved modification of the bisulphite based sequencing method. Our strategy is to perform the bisulphite treatment and subsequent PCR steps on material embedded into agarose beads. This prevents loss of DNA during the experimental procedure and ensures an optimal bisulphite reactivity by maintaining the DNA in the single stranded form. The modification improves previously published protocols in that it facilitates the handling of probes and reproducibly reaches a very high level of sensitivity.
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Cited by
Chronic high-fat diet in fathers programs β-cell dysfunction in female rat offspring
Ng, S.-F., Lin, R. C. Y., Laybutt, D. R., Barres, R., Owens, J. A., & Morris, M. J. (2010) · Nature · doi:10.1038/nature09491 · Cited by 1,327
Active demethylation of the paternal genome in the mouse zygote
Oswald, J., Engemann, S., Lane, N., Mayer, W., Olek, A., Fundele, R., Dean, W., Reik, W., & Walter, J. (2000) · Current Biology · doi:10.1016/s0960-9822(00)00448-6 · Cited by 981
Self-citationAcute Exercise Remodels Promoter Methylation in Human Skeletal Muscle
Barrès, R., Yan, J., Egan, B., Treebak, J. T., Rasmussen, M., Fritz, T., Caidahl, K., Krook, A., O'Gorman, D. J., & Zierath, J. R. (2012) · Cell Metabolism · doi:10.1016/j.cmet.2012.01.001 · Cited by 965
“Bisulfite treatment was performed as described (Olek et al., 1996), with the following adaptations: 1 mg of genomic DNA was embedded in a 2% Low Melting Point Agarose solution and 10 beads were formed.”
MethodNon-CpG methylation of the PGC-1α promoter through DNMT3B controls mitochondrial density.
Barrès, R., Osler, M. E., Yan, J., Rune, A., Fritz, T., Caidahl, K., Krook, A., & Zierath, J. R. (2009) · Cell Metabolism · doi:10.1016/j.cmet.2009.07.011 · Cited by 602
DNA Methylation Analysis by Pyrosequencing
Busato, F., & Tost, J. (2014) · Encyclopedia of Metagenomics · doi:10.1007/978-1-4614-6418-1_799-1 · Cited by 499
Metadata
- Authors
- Olek, A., Oswald, J., & Walter, J.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1996-12-01
- Volume
- 24
- Issue
- 24
- Pages
- 5064-5066
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2064838548
- Cited by
- 376
- References
- 0
- FWCI
- 1.61
- Type
- journal article
- Language
- en
- Open access
- green
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- CoChat. (2026). Citation check: A Modified and Improved Method for Bisulphite Based Cytosine Methylation Analysis. https://app.cochat.ai/papers/10.1093/nar/24.24.5064
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- Olek, A., Oswald, J., & Walter, J. (1996). A Modified and Improved Method for Bisulphite Based Cytosine Methylation Analysis. Nucleic Acids Research. https://doi.org/10.1093/nar/24.24.5064
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