GSM5939637: Input DNA; Oryza sativa; ChIP-Seq
Total DNA of Nipponbare and purified GST-APIP5 and GST were used for ChIP assays. DNA was prepared from 1-month-old rice leaves using CATB method. The total DNA was sheared into 100-500bp fragments using ultrasonic crusher. The recombinant proteins GST-APIP5 and GST were expressed in Escherichia coli, and were purified with Glutathione Sepharose 4B (GE Healthcare). GST-APIP5 or GST was incubated with an equal amount of DNA fragments for 2h, respectively. The incubation buffer includes: 50mM Tris (pH 7.0), 1mM EDTA, 100mM KCl, 5% Glycerol, 0.1% Triton X-100, 1mM DTT. After co-incubation, the beads were washed 5-10 times using incubation buffer. The precipitated DNA fragments were recovered eith CTAB and dissolved in sterile water for constructing Illumina sequencing library. Libraries were prepared according to Illumina's instructions.