ORGANIC
Occupied Regions of Genomes from Affinity-Purified Naturally Isolated Chromatin
ORGANIC is a gentle protocol that avoids crosslinking and sonication (Kasinathan et al., 2014). The method is a combination of MNase-Seq and native ChIP that provides accurate maps of chromatin-occupied regions in complex eukaryotic genomes.
Advantages:
- Avoids sonication bias (Teytelman et al., 2009)
- Avoids crosslinking artifacts (Zentner et al., 2014)
Disadvantages:
- Potentially poor solubilization of proteins (Zentner et al., 2014)
- MNase sequence bias (Kensche et al., 2016)
- Nuclei isolation may introduce artifacts
- Not replicated in other laboratories
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Zentner G. E. and Henikoff S. High-resolution digital profiling of the epigenome. Nat Rev Genet. 2014;15:814-827
References:
Kasinathan S., Orsi G. A., Zentner G. E., Ahmad K. and Henikoff S. High-resolution mapping of transcription factor binding sites on native chromatin. Nat Methods. 2014;11:203-209
Zentner G. E. and Henikoff S. Mot1 redistributes TBP from TATA-containing to TATA-less promoters. Mol Cell Biol. 2013;33:4996-5004