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PLoS One ; 14(11): e0225801, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31765427

RESUMEN

Chromatin immunoprecipitation coupled with next-generation sequencing (ChIP-seq) has revolutionized our understanding of chromatin-related biological processes. The method, however, requires thousands of cells and has therefore limited applications in situations where cell numbers are limited. Here we describe a novel method called Restriction Assisted Tagmentation Chromatin Immunoprecipitation (RAT-ChIP) that enables global histone modification profiling from as few as 100 cells. The method is simple, cost-effective and takes a single day to complete. We demonstrate the sensitivity of the method by deriving the first genome-wide maps of histone H3K4me3 and H3K27me3 modifications of inner cell mass and trophectoderm of bovine blastocyst stage embryos.


Asunto(s)
Masa Celular Interna del Blastocisto/metabolismo , Inmunoprecipitación de Cromatina , Histonas/metabolismo , Trofoblastos/metabolismo , Animales , Blastocisto/citología , Blastocisto/metabolismo , Bovinos , Línea Celular , Cromatina/química , Cromatina/metabolismo , Fertilización In Vitro , Genoma , Secuenciación de Nucleótidos de Alto Rendimiento , Histonas/genética , Humanos , Oocitos/citología , Procesamiento Proteico-Postraduccional , Análisis de Secuencia de ADN
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