GSM5014385: CLSY3-3xFLAG_Fl_IP; Arabidopsis thaliana; ChIP-Seq
~2.0 g of un-opened flower buds (stage 12 and younger) was collected, ground in liquid nitrogen, and in vitro crosslinked with 1% formaldehyde (20 minutes at RT; Sigma, Cat# F8775). The chromatin was then fragmented to ~500bp by sonication, incubated with anti-Flag M2 Magnetic beads (4 °C for 2 h; Sigma, Cat# M8823), washed 5 times and then eluted with 3xFLAG peptide ([0.1 mg/mL]; Sigma, Cat# F4799). The crosslinking was reversed overnight at 65 °C overnight and purified DNA (Thermo Scientific, Cat# 17908) was used to generate libraries with the NEBNext Ultra II DNA Library Prep Kit (New England Biolabs, Cat# 7645). The libraries were sequenced (single end 50 bp, SE50) on a HiSeq 2500 machine (Illumina).