GSM5195676: hen1fbw2_3 (AGO1-IP RNA); Arabidopsis thaliana; ncRNA-Seq
RNA was extracted in Tri-Reagent according to the manufacturer's protocol, and aqueous phase was left in 1 volume of isopropanol over-night at -20°C, precipitated 30 minutes at 16000g (4°C). Pellets were rinsed in 1 ml 70% ethanol and centrifuged an additional 10 minutes. For AGO1-loaded small RNA samples, IPs were performed as described above from 500mg of 1-week-old Arabidopsis seedlings grown on MS-agar plates. AGO1-loaded small RNA were then extracted by adding Tri-Reagent directly on the magnetic beads and extraction of RNA was then performed according to the manufacturer's instructions. Small RNA libraries were constructed using Real-Seq-AC kit (RealSeq®-AC, USA) according to manufacturer's instructions with 500ng as starting material. Libraries were sequenced using Illumina Next-Seq 500 technology at University of Delaware (Delaware, USA).