Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
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
After thawing frozen PBMC samples at 37uC, total RNA was immediately extracted using a Maxwell16 LEV simplyRNA Blood Kit on a Maxwell16 Instrument (Promega) according to the manufacturer’s instructions. A 4-mL aliquot of each sample of extracted total RNA was used for RNA quality control assessments, while the remaining RNA sample was stored at 2 80uC until use. The RNA yield was measured using a Qubit RNA Assay Kit on a Qubit2.0 Fluorometer (Life Technologies). RNA purity was estimated by measuring the ratio of absorbance at 260 and 280 nm (A260/A280) using a Nanodrop 2000 UV-Vis Spectrophotometer (Thermo Fisher Scientific). The RNA integrity number (RIN) was assessed using an RNA 6000 Nano Kit on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s instructions
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