MNase-seq of sorghum: root
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 nuclei was resuspended with Micrococcal Nuclease (MNase) buffer, and then BSA and MNase were added and then digested at 37 for 5 min, 10 min, 15 min. The reaction was stopped by adding EGTA. RNase and 20ul 5M NaCl were used for removing RNA and de-crosslinking respectively by incubating at 65 overnight. DNA was purified and separated by 2% agarose gel electrophoresis. Around 200bp DNA fragments generated by digestion of a proper time were recovered with gel extraction kit.