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
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