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AHT-ChIP-seq: a completely automated robotic protocol for high-throughput chromatin immunoprecipitation.
Genome Biol ; 14(11): R124, 2013 Nov 07.
Article em En | MEDLINE | ID: mdl-24200198
ABSTRACT
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Robótica / Imunoprecipitação da Cromatina / Sequenciamento de Nucleotídeos em Larga Escala Tipo de estudo: Prognostic_studies / Qualitative_research Limite: Animals / Humans / Male Idioma: En Ano de publicação: 2013 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Robótica / Imunoprecipitação da Cromatina / Sequenciamento de Nucleotídeos em Larga Escala Tipo de estudo: Prognostic_studies / Qualitative_research Limite: Animals / Humans / Male Idioma: En Ano de publicação: 2013 Tipo de documento: Article