alias: GSM2809097
title: GSM2809097: KO-cytoRNA-2.0_1st 2.0 ng/ul RNase A; Homo sapiens; OTHER
library_construction_protocol: cytoplasmic RNA was purified with hypotonic buffer, followed by douncing 50 times. RNA was purified using Qiagen RNeasy Mini Kit. RNAs were first incubated with recombinant  WT MDA5 proteins, then challenged by 2ng/ul of RNaseA. The reaction was terminated by TRIzol. RNA was recovered and rRNA was depleted using NEBNext rRNA depletion kit. cDNA library was constructed using Clontech Smarter Stranded RNA-seq Kit. Mock-treated RNAs (cytoRNA-0.0) were fragmented by heating at 94C for 3min, RNaseA-digested RNA (cytoRNA-2.0) was fragmented by heating at 94C for 1min without First-strand Buffer. PCR cycles for cytoRNA-0.0 was 15, for cytoRNA-2.0 were 18. The Ampure Beads:DNA ratio was 0.8:1 (volume:volume) for DNA purification.
identifiers.namespace: GEO
identifiers.id: GSM2809097