GSM5745372: DCL1ci-IP_rep2; Arabidopsis thaliana; RIP-Seq
Three independent pools of pDCL1:DCL1ci-HA/dcl1-7 and three independent pools of dcl1-7 (control plants) were fixed in formaldehyde 1%, frozen and ground in liquid nitrogen. Supernatants were sonicated, precleared and incubated with Pierce™ Anti-DYKDDDDK Magnetic Agarose (Thermo Scientific™). The beads were removed magnetically, washed four times and reversal crosslinked in NaCl 200mM and Proteinase K. RNA was then phenol-extracted and precipitated in isopropanol and glycogen, washed in ethanol 80% and re-suspended in RNase-free water. Extracted RNA was processed into sequencing libraries using adapted Illumina protocols and sequenced at Fasteris (http://www.fasteris.com, Switzerland) using the Illumina HiSeq sequencer