GSM2650288: NRPD1-3xFLAG,clsy1-7_IP_1; Arabidopsis thaliana; ChIP-Seq
2.0g of un-opened flower buds (stage 12 and younger) for each genotype were collected and ground to a fine powder in liquid nitrogen and crosslinked in 1% formaldehyde (Sigma, Cat# F8775) for 20min at room temperature with slow rotation. The chromatin was then fragmented to ~500bp by sonication and the lysate was incubated with the anti-Flag M2 Magnetic beads (Sigma, Cat# M8823) at 4°C for 2h. The beads were washed 5 times, for 5min at 4°C and then the crosslinking was reversed by incubation at 65°C overnight and further purified using the phenol:Chloroform:Isoamyl Alcohol kit (Thermo scientific, Cat# 17908). The resulting DNA then was used for library preparation with NEBNext Ultra II DNA Library Prep Kit (NEB, Cat# 7645). The ChIP libraries were sequenced in SE50 mode on a HiSeq2500 machine.
GEO Accession
GSM2650288