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

被引：24

作者：

Aldridge, Sarah ^{[1
]}

Watt, Stephen ^{[1
]}

Quail, Michael A. ^{[2
]}

Rayner, Tim ^{[1
]}

Lukk, Margus ^{[1
]}

Bimson, Michael F. ^{[3
]}

Gaffney, Daniel ^{[2
]}

Odom, Duncan T. ^{[1
,2
]}

机构：

[1] Univ Cambridge, Canc Res UK Cambridge Inst, Li Ka Shing Ctr, Cambridge CB2 0RE, England

[2] Wellcome Trust Sanger Inst, Cambridge CB10 1HH, England

[3] Agilent Technol UK Ltd, Kidlington OX5 1QU, Oxon, England

来源：

GENOME BIOLOGY | 2013年 / 14卷 / 11期

基金：

欧洲研究理事会; 英国惠康基金;

关键词：

TRANSCRIPTION FACTOR-BINDING; HUMAN GENOME; DNA; METHYLATIONS; NESTEDMICA; SEQUENCE; REVEALS;

D O I：

10.1186/gb-2013-14-11-r124

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页数：12

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