Protocol: methodology for chromatin immunoprecipitation (ChIP) in Chlamydomonas reinhardtii

被引：0
|
作者：
Daniela Strenkert
Stefan Schmollinger
Michael Schroda
机构：
[1] Max-Planck-Institut für Molekulare Pflanzenphysiologie,
来源：
Plant Methods | / 7卷
关键词：
Chromatin immunoprecipitation; heat shock; formaldehyde crosslinking; real-time PCR; nucleosome occupancy; histone modification;
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学科分类号：
摘要：
We report on a detailed chromatin immunoprecipitation (ChIP) protocol for the unicellular green alga Chlamydomonas reinhardtii. The protocol is suitable for the analysis of nucleosome occupancy, histone modifications and transcription factor binding sites at the level of mononucleosomes for targeted and genome-wide studies. We describe the optimization of conditions for crosslinking, chromatin fragmentation and antibody titer determination and provide recommendations and an example for the normalization of ChIP results as determined by real-time PCR.
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