Qubit RNA HS Assay Kit

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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	The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay

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
The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay

Publication protocol

mRNA isolation for LC-MS/MS: total RNA was isolated from wild-type or transiently transfected cells with TRIzol reagent (Invitrogen). mRNA was extracted using Dynabeads® mRNA DIRECT kit (Ambion) followed by further removal of remaining rRNA using RiboMinus Eukaryote Kit v2 (Ambion). mRNA concentration was measured by Qubit® RNA HS Assay Kit with Qubit® 2.0 fluorometer.

RNA isolation for RT-qPCR: we followed the instruction of Direct-zol™ RNA Miniprep kit (Zymo Research) or RNeasy Miniprep kit (Qiagen) both with an in-column DNase I digestion step. For RT-qPCR following polysome profiling, RNA was extracted according to the manufacturers manual of TRizol® reagent (added 1:1), and contaminate DNA was removed using RNA clean and concentrator-25 (Zymo Research) with additional in-column DNase I digestion.

Ethanol precipitation: to the RNA solution being purified or concentrated, 1/10 volume of 3 M NaOAc, pH 5.5, 1 μl GlycoBlue™ coprecipitant (15 mg/ml, Ambion), and 2.7 volume of 100% ethanol were added. The mixture was stored at −20 °C overnight, and then centrifuged at 15 000 × g for 15 min. After removing the supernatant, the pellet was washed twice with 1 ml 75% ethanol, and dissolved in the appropriate amount of RNase-free water as indicated.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Qubit RNA HS Assay Kit below.

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