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Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells
Authors:Geoffrey Berguet  Jan Hendrickx  Celine Sabatel  Miklos Laczik  Sharon Squazzo  Ignacio Mazon Pelaez  Rini Saxena  Helene Pendeville  Dominique Poncelet
Institution:1.Diagenode S.A.;2.Diagenode Inc.
Abstract:Chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq) is a technique of choice for studying protein-DNA interactions. ChIP-seq has been used for mapping protein-DNA interactions and allocating histones modifications. The procedure is tedious and time consuming, and one of the major limitations is the requirement for high amounts of starting material, usually millions of cells. Automation of chromatin immunoprecipitation assays is possible when the procedure is based on the use of magnetic beads. Successful automated protocols of chromatin immunoprecipitation and library preparation have been specifically designed on a commercially available robotic liquid handling system dedicated mainly to automate epigenetic assays. First, validation of automated ChIP-seq assays using antibodies directed against various histone modifications was shown, followed by optimization of the automated protocols to perform chromatin immunoprecipitation and library preparation starting with low cell numbers. The goal of these experiments is to provide a valuable tool for future epigenetic analysis of specific cell types, sub-populations, and biopsy samples.
Keywords:Molecular Biology  Issue 94  Automation  chromatin immunoprecipitation  low DNA amounts  histone antibodies  sequencing  library preparation
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