AHT-ChIP-seq: a completely automated robotic protocol for high-throughput chromatin immunoprecipitation
- Equal contributors
1 University of Cambridge, Cancer Research UK - Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge, CB2 0RE, UK
2 Wellcome Trust Sanger Institute, Hinxton, Cambridge, CB10 1HH, UK
3 Agilent Technologies UK Limited, Mead Road, Yarnton, Kidlington, Oxfordshire, OX5 1QU, United Kingdom
Genome Biology 2013, 14:R124 doi:10.1186/gb-2013-14-11-r124Published: 7 November 2013
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.