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AHT-ChIP-seq: a completely automated robotic protocol for high-throughput chromatin immunoprecipitation

ChIP-seq is an established manually-performed method for identifying DNA-protein interactions genome-wide. Here, we describe a protocol for automated high-throughput (AHT) ChIP-seq. To demonstrate the quality of data obtained using AHT-ChIP-seq, we applied it to five proteins in mouse livers using a...

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Detalles Bibliográficos
Autores principales: Aldridge, Sarah, Watt, Stephen, Quail, Michael A, Rayner, Tim, Lukk, Margus, Bimson, Michael F, Gaffney, Daniel, Odom, Duncan T
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4053851/
https://www.ncbi.nlm.nih.gov/pubmed/24200198
http://dx.doi.org/10.1186/gb-2013-14-11-r124
Descripción
Sumario:ChIP-seq is an established manually-performed method for identifying DNA-protein interactions genome-wide. Here, we describe a protocol for automated high-throughput (AHT) ChIP-seq. To demonstrate the quality of data obtained using AHT-ChIP-seq, we applied it to five proteins in mouse livers using a single 96-well plate, demonstrating an extremely high degree of qualitative and quantitative reproducibility among biological and technical replicates. We estimated the optimum and minimum recommended cell numbers required to perform AHT-ChIP-seq by running an additional plate using HepG2 and MCF7 cells. With this protocol, commercially available robotics can perform four hundred experiments in five days.