The DeNovix® RNA Assay enables the accurate detection of RNA samples with a standard detection range of 5 ng – 1500 ng total mass in 200 µL volumes. This equates to sample concentrations of 250 pg/µL – 1500 ng/µL when using between 1 and 20 µL sample volumes in 200 µL assay volumes. The assay is linear for sample concentrations as high as 1500 ng/µL when adjusting volumes to 1 µL of sample into 199 µL of working reagent.
The DeNovix RNA Assay is available for use on Qubit® 2.0, 3.0, and Qubit® 4 Fluorometers. The assay parameter files are available on the DeNovix website.
Three assay sizes are available. The volume of components in each kit are sufficient for 1000, 250 and 50 (evaluation size) assays respectively. Kit components are shown in Table 1.Table 1: DeNovix RNA Assay Kit Components
|DeNovix RNA Quantitation Dye (200x)||1 mL||250 µL||50 µL|
|DeNovix RNA Assay Buffer||250 mL||62.5 mL||12.5 mL|
|100 ng/µL RNA Standard (mammalian cell)||4 x 400 µL||400 µL||0.1 mL|
|0 ng/µL RNA Standard||2 mL||0.5 mL||0.5 mL|
Installing DeNovix Assays on a Qubit® Fluorometer
For a Qubit® 2.0 Fluorometer:
For Qubit® 3.0 and Qubit® 4 Fluorometers:
Once the assay file is uploaded to the Qubit® Fluorometer, simply select the DeNovix RNA Assay to measure RNA samples.
Measurement Data For Qubit® 2.0
Three replicates of RNA samples were measured on a Qubit® 2.0 and a DeNovix DS-11 FX Fluorometer using the DeNovix RNA Assay. Each sample was prepared by adding between 1-20 µL stock to 199-180 µL working solution respectively, depending on the total mass limits of the assay. The data is presented in graphical and tabular form across the assay dynamic range below.Table 2: The data presented demonstrate the measurement range of the DeNovix RNA Fluorescence Assay measured on a DeNovix DS-11 FX Fluorometer and Qubit® 2.0.
|Expected RNA||Qubit® 2.0||DS-11 FX|
|1500||Measured Out of Range||1530.39||22.029|
Measurement Data For Qubit® 3.0 / 4
Three replicates of RNA samples were measured on a Qubit® 3.0. a Qubit® 4, and a DeNovix DS-11 FX Fluorometer using the DeNovix RNA Assay. The data is presented in graphical and tabular form across the assay dynamic range below.
|Expected RNA||Qubit® 3.0||Qubit® 4||DS-11 FX|
|1500||Measured Out of Range||Measured Out of Range||1530.39||22.029|
The DeNovix RNA Assay enables specific, highly sensitive RNA quantitation across a wide dynamic range. The assay is available for use on a Qubit® Fluorometer through the simple application of downloadable assay files.
Qubit® is a registered trademark of Thermo Fisher Scientific and its subsidiaries.
Revised 19 Oct 2020