Qubit RNA HS Assay Kit

Protocol tips

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The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay	Dilute Qubit® RNA HS Reagent 1:200 in Qubit® RNA HS Buffer

Upstream tips
The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay
Protocol tips
Dilute Qubit® RNA HS Reagent 1:200 in Qubit® RNA HS Buffer

Publication protocol

RNA samples were quantitated using the Qubit HS RNA kit (Thermo Fisher Scientific) with a Qubit 3.0 Fluorometer (Thermo Fisher Scientific), following manufacturer’s instructions. The 260/280 and 260/230 ratios of absorbance values were used to assess the purity of RNA using a Nanodrop ND-1000 spectrophotometer (Thermo Fisher Scientific). A 260/280 ratio of ~ 2.0 and 260/230 ratio in the range of 2.0–2.2 was accepted as “pure” for RNA. Lower ratios may indicate the presence of protein, phenol, EDTA, carbohydrates or other contaminants that absorb at or near 260, 230 or 280 nm. The miRNA content was quantitated using a Qubit microRNA assay kit (Thermo Fisher Scientific) with a Qubit 3.0 Fluorometer, according to manufacturer’s recommendations. The integrity of the RNA was determined for all samples using an Agilent Bioanalyzer 2100 (Agilent Technologies, Diegem, Belgium) with an Agilent RNA 6000 kit (Agilent Technologies). RNA Integrity Numbers (RINs) were used to evaluate the integrity of the RNA samples with > 7.0 considered intact and < 7.0 considered degraded. The quality assessment of RNA samples is provided in Additional file 1: Figure S2, Additional file 2: Figure S3, Additional file 3: Figure S4.

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