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

Sarah Aldridge, Stephen Watt, Michael A. Quail, Tim F. Rayner, Margus Lukk, Michael F Bimson, Daniel J. Gaffney, Duncan T. Odom

Year
2013
Citations
45
Access
Open access

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.

Keywords

Protocol (science)Computer scienceThroughputChromatin immunoprecipitationChipImmunoprecipitationEmbedded systemComputational biologyArtificial intelligenceBiology

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