GSM7419615: mc-HMGA_input_rep2; Arabidopsis thaliana; ChIP-Seq
5 g leaves were cross-linked in 1% formaldehyde for 10 min. Then stopped by adding 0.125 M glycine and washed by PBS. The nuclei were extrected by Honda buffer and washed 3 times by NRBT. Then the nuclei were sonicated 4 times for 10 min (30 s on/off intervals) using the Bioruptor® Pico. The sonicated nuclei were centrifuged at 15000rpm for 15 min, and the supernatant was transferred to new tubes and diluted 10 times with ChIP dilution buffer (1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8.0, 167 mM NaCl). 50 μL diluted supernatant was preserved as input. Then 2 μg anti-GFP antibody (Abcam, ab290), anti-mCherry antibody (Abcam, ab213511), or anti-H3K9me2 antibody (Abcam, ab1220) was added into supernatant and rotate at 4 °C overnight. 50 μL washed Protein G Dynabeads (Invitrogen, 10004D) was added to the sample. After rotating for 4 h, the beads were washed sequentially with Low Salt Buffer, High Salt Buffer, LiCl Buffer, TE Buffer (each buffer for a quick wash, and a 10 min wash). Then add 100 μL 10% Chelex resin (Biorad, 1422842) to elute immune complexes by incubating at 95 °