Name: GSM8268680
Instrument: Illumina NovaSeq 6000
Strategy: ncRNA-Seq
Source: OTHER
Selection: size fractionation
Layout: PAIRED
Construction protocol: To extract nucleic acids co-purified with HrAgo1, 2 nmoles of purified protein was incubated with 250 µg/ml proteinaseK (Thermo Scientific) for 4h at 65C. Next, phenol:chloroform:IAA 25:24:1 pH 7.9 (Invitrogen) was added in a 1:1 ratio. The sample was vortexed and centrifuged at 16000 x g in a table top centrifuge for 10 min. The upper layer containing the nucleic acids was transferred to a clean tube and the nucleic acids were precipitated through ethanol precipitation. To this end, 99% cold ethanol and 3 M sodium acetate pH 5.2 were added to the sample in a 2:1 and 1:9 ratio, respectively. The sample was incubated overnight at -80°C, after which it was centrifuged at 16000 x g in a table top centrifuge for 1 h. The pellet was washed with 70% ethanol and subsequently dissolved in nuclease-free water. The small RNAseq library was constructed by GenomeScan (Leiden, NL) using standard small RNA library preparation protocols