Instrument: Illumina HiSeq 2000
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Fixation of the plant material and ChIP was performed as described in Gendrel, et al. 2005, (Nature Methods, 2(3), 213–8). Nuclear extracts were split in two and incubated each with 20 µl of GFP-Trap_A (ChromoTek) for 4 h at 5°C for immunoprecipitation. MinElute Reaction Cleanup Kit columns (Qiagen) were used for purification of the DNA fragments. Sequencing libraries were generated using pooled DNA from each 7 to 10 individual ChIP preparations of either RGA-GFP or WT material. Libraries were prepared for Illumina Sequencing using the NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina (E6240) with the NEBNext Multiplex Oligos for Illumina (E7335 and E7500). Libraries were quantified using the Qubit Fluorometer from Invitrogen, and their size determined using the DNA 1000 Assay on the BioAnalyzer. Sequencing was performed on the Illumina HiSeq 2000 Instrument 50 base SE (plus index read).