GSM6598256: MH_seedling_H3K27me3_ChIA_Rep2; Oryza sativa Indica Group; ChIA-PET
Rice tissues were dual cross-linked with 1% formaldehyde (SIGMA, F8775) for 10 min and 1.5 mM ethylene glycol bis (succinimidylsuccinate) (Thermo Fisher Scientific, 21565) for 30 min at 25 ℃, after which glycine was added to a final concentration of 0.2 M to stop the cross-linking reaction. pl-DNA was end-repaired, A-tailed, and proximity ligated using a single biotinylated bridge-linker (Forward strand: 5′-[5Phos]CGCGATATC/iBIOdT/TATCTGACT-3′, Reverse strand: 5′-[5Phos]GTCAGATAAGATATCGCGT-3′). Proximity ligated protein-DNA complexes were eluted and reverse cross-linked (similar to that in eChIP-Seq). Tn5 transposase (VAHTS， TD501) was used to prepare ChIA-PET libraries according to the operation manual, and the libraries were sequenced (paired end 150 bp) using an Illumina HiSeq X-Ten (AnnoRoad Company, Beijing, China).