Instrument: Illumina HiSeq 2500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Please see accompanying publication for detailed protocol. Briefly, ChIP-seq experiments for PIF4 and CRY2 were performed as described (Kaufmann et al., 2010) using 5 day-old seedlings harboring either a PIF4-Flash (Thines et al., 2014) or UBQ10pro::Flash-CRY2 transgene. A monoclonal anti-FLAG antibody (Sigma) was used in all experiments. The CRY2 ChIP experiment was performed with two modifications. Instead of 2g starting material that we used to precipitate PIF4, 18g starting material was used for CRY2. Moreover, we conducted a dual cross-linking procedure for CRY2 using the additional cross-linker ethylene glycol bis-succinimidylsuccinate. ChIP DNA was used to generate sequencing libraries as per Illumina protocol (Illumina, CA). Libraries were prepared according to Illumina's instructions accompanying TruSeq DNA sample prep kit and sequenced by Illumina HiSeq 2500 according to the manufacturer's instructions.