Post-conversion targeted capture of modified cytosines in mammalian and plant genomes

We present a capture-based approach for bisulfite-converted DNA that allows interrogation of pre-defined genomic locations, allowing quantitative and qualitative assessments of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) at CG dinucleotides and in non-CG contexts (CHG, CHH) in mammalia...

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Detalles Bibliográficos
Autores principales: Li, Qing, Suzuki, Masako, Wendt, Jennifer, Patterson, Nicole, Eichten, Steven R., Hermanson, Peter J., Green, Dawn, Jeddeloh, Jeffrey, Richmond, Todd, Rosenbaum, Heidi, Burgess, Daniel, Springer, Nathan M., Greally, John M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2015
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4499119/
https://www.ncbi.nlm.nih.gov/pubmed/25813045
http://dx.doi.org/10.1093/nar/gkv244
Descripción
Sumario:We present a capture-based approach for bisulfite-converted DNA that allows interrogation of pre-defined genomic locations, allowing quantitative and qualitative assessments of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) at CG dinucleotides and in non-CG contexts (CHG, CHH) in mammalian and plant genomes. We show the technique works robustly and reproducibly using as little as 500 ng of starting DNA, with results correlating well with whole genome bisulfite sequencing data, and demonstrate that human DNA can be tested in samples contaminated with microbial DNA. This targeting approach will allow cell type-specific designs to maximize the value of 5mC and 5hmC sequencing.

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