Mapping the distribution of chromatin proteins by ChIP on chip.
The ChIP on chip method combines chromatin immunoprecipitation (ChIP) with hybridization on DNA microarrays (chip). The ChIP technique allows one to obtain a DNA sample enriched in sequences bound by transcription factors or chromatin-associated proteins. Usually, ChIP is used to test whether specific candidate sequences are bound by a transcription factor, but microarrays are a powerful tool that allows testing large pools of sequences at once. This chapter presents the pipeline of a ChIP on chip method that can be applied to map the binding sites of chromatin-associated proteins along Drosophila chromosomes at different developmental stages. This chapter provides protocols for ChIP, for quality control tests of ChIP samples, for microarray design, for hybridization of the ChIP samples onto microarrays, and for initial analysis of the data. In addition, this chapter discusses the most important steps in each of the protocols as well as the importance of bioinformatic analysis in order to extract valuable biological information from the data sets.
Nègre
Nicolas
N
Chromatin and Cell Biology Lab, Institute of Human Genetics- CNRS, Montpellier, Cedex 05, France.
Lavrov
Sergey
S
Hennetin
Jérôme
J
Bellis
Michel
M
Cavalli
Giacomo
G
D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones