title: AHT-ChIP-seq: a completely automated robotic protocol for high-throughput chromatin immunoprecipitation.
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.
authors.forename: Sarah
authors.initials: S
authors.lastname: Aldridge
authors.forename: Stephen
authors.initials: S
authors.lastname: Watt
authors.forename: Michael A
authors.initials: MA
authors.lastname: Quail
authors.forename: Tim
authors.initials: T
authors.lastname: Rayner
authors.forename: Margus
authors.initials: M
authors.lastname: Lukk
authors.forename: Michael F
authors.initials: MF
authors.lastname: Bimson
authors.forename: Daniel
authors.initials: D
authors.lastname: Gaffney
authors.forename: Duncan T
authors.initials: DT
authors.lastname: Odom
mesh_terms: D000818:Animals; D047369:Chromatin Immunoprecipitation / Q000295:instrumentation / Q000379:methods*; D056945:Hep G2 Cells; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008099:Liver / Q000378:metabolism; D061986:MCF-7 Cells; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation / Q000379:methods*; D017422:Sequence Analysis, DNA