ChIP-seq of sorghum: root under PEG
Samples were crosslinked with 1%(v/v) formaldehyde under vacuum for 30min and then stopped by adding 2M glycine for another 5min. The nuclei were extracted using extraction buffer I, extraction buffer II and extraction buffer III, and split in nuclei lysis buffer. The chromatin DNA was broken into 200-500bp fragments by sonication. The antibodies were incubated with Protein A agarose in ChIP dilution buffer before mixed with sonicated chromatin. The immunoprecipitated agarose beads were washed with low salt buffer, high salt buffer, LiCl buffer and TE buffer, and then eluted with elution buffer. After de-crosslinking, The eluate was treated with Protease K and RNase H and DNA was purified.