TN 169 Thermo Fisher Qubit™ RNA Assay Performance Data

RNA Quantification Assays

The Thermo Fisher Qubit™ RNA HS assay enables selective quantitation in the standard detection range of 5 – 100 ng total mass of RNA when using 1 – 20 μL in 200 μL assay volume. This equates to measuring RNA initial sample concentrations of 250 pg/μL – 100 ng/μL when using 1 – 20 μL of sample in 200 μL assay volume.

The standard detection range of the Thermo Fisher Qubit™ RNA BR assay is 20 – 1000 ng total mass when using 1 – 20 μL in 200 μL assay volume. This equates to measuring RNA initial sample concentrations of 1 – 1000 ng/μL when using 1 – 20 μL of sample in 200 μL assay volume.

The Thermo Fisher Qubit™ RNA HS assay kit (Thermo Fisher Scientific cat #Q32852) and the Qubit™ RNA BR assay kit (Thermo Fisher Scientific cat #Q10210) were used to perform both assays. Each kit has a mix-and-measure protocol and includes buffer, reagent and two standards. Samples were measured in thin-walled, clear UV-transparent 0.5 mL PCR tubes (DeNovix cat #TUBE-PCR-0.5-500).

Performance data for Thermo Fisher Qubit™ RNA quantification assays were obtained on a Thermo Fisher Qubit™ 3.0 fluorometer and a DeNovix QFX Fluorometer. Each assay was prepared as described in the manufacturer’s protocol. Samples were mixed and incubated at room temperature for 5 minutes. Three replicate measurements were taken for each sample.

A series of dilutions was gravimetrically prepared in HPLC grade water from the Thermo Fisher Qubit™ HS and BR assay standards. The samples for the HS assay were prepared from the 10 ng/μL standard, and the samples for the BR assay were diluted from the 100 ng/μL standard. Each sample was prepared to fall within the total mass limits of the respective assay. For each assay, 1 – 20 μL of each prepared sample was added to 180 – 190 μL working solution. Initial sample concentrations were between 0.25 and 10 ng/μL for the HS assay and between 1 and 100 ng/μL for the BR assay.