GSM1624648: GL×93-11_H3K27me3 _ChIP-seq; Oryza sativa; ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed using antibodies against H3K27me3 (Abcam, Cat. #ab6002, Cambridge, MA 02139-1517, USA) and H3K36me3 (Abcam, Cat. #ab9050). DNA was extracted using an equal volume phenol:chloroform:isoamyl alcohol and briefly vortexed; DNA was precipitated with a 2.5-volume of 100% EtOH containing 0.3 M sodium acetate and 2 μl of glycogen (20 mg ml–1) at a pH of 5.2. The resulting ChIP DNA was used to generate Illumina sequencing libraries according to the manufacturer’s protocol: suitable fragments of approximately 200 bp were selected as templates for amplification after incubation at 98°C for 30 s for denaturation, followed by 15 cycles of 98°C for 10 s, 65°C for 30 s, 72°C for 30 s and 72°C for 5 min. The samples were purified using the QIAquick PCR purification kit (Qiagen, Valencia,CA USA) as described in the manufacturer’s protocol.