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キーワード:
chromatin immunoprecipitation
chlamydomonas reinhardtii
heat shock
formaldehyde crosslinking
real-time pcr
nucleosome occupancy
histone modification
hsp70a
rbcs2
cyc6
carbon-concentrating mechanism
protein DNA interactions
heat-shock
gene
formaldehyde
acetylation
regulator
regions
core
要旨:
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.