Question 1

What is the quantification range?

Accepted Answer

The target mass range is 5–100 ng in the assay. The stated initial concentration range is 0.5–100 ng/µL, depending on sample input. Initial sample concentration is measured before dilution; target mass equals initial concentration multiplied by the sample volume. These are different quantities.

Question 2

How do I prepare the working solution?

Accepted Answer

Prepare fresh working solution by mixing 1 volume of fluorescent reagent with 199 volumes of the supplied buffer (1:200 final dilution).

Question 3

Can I use this kit with Qubit fluorometers?

Accepted Answer

The supplier protocol supports use with the matching Qubit assay program. Use this kit's own standards, the specified sample volume and the correct consumables for the Qubit model. Compatibility does not imply Thermo Fisher endorsement or identical performance. For the 1X assay on older Qubit models, verify that the appropriate assay program is installed.

Question 4

Which Biofargo fluorometers support this assay?

Accepted Answer

X1 and X3 both list RNA HS and RNA BR programs.

Question 5

How much sample is required?

Accepted Answer

Add 1–20 µL of sample and adjust the working solution to a final volume of 200 µL. Choose the input volume so the target mass lies within the assay range. Dilute samples that exceed the upper limit.

Question 6

Which assay tubes should I use?

Accepted Answer

Use clear, thin-wall 0.5 mL assay tubes for Biofargo X1 and Qubit 2/3/4. Use the instrument-specified clear 0.2 mL tubes or eight-tube strips for Biofargo X3; Qubit Flex requires its specified 0.2 mL assay tube strips. Keep the total assay volume at 200 µL.

Question 7

How long does the assay take?

Accepted Answer

Incubate for 2 minutes at 20–25 °C, protected from light, after mixing. This is the incubation time, not the total preparation and measurement time. The signal remains stable for up to 3 hours when protected from light.

Question 8

Do I need new standards?

Accepted Answer

Use the two standards supplied with this kit and the matching assay program. A new calibration for each run is recommended. Do not reuse a calibration generated with a different kit. For a plate reader, prepare a dilution series and fit a standard curve.

Question 9

Does the reaction count equal the number of unknown samples?

Accepted Answer

The listed count is the nominal assay format. Standards, controls, repeat measurements and handling losses reduce the number of unknown samples that can be measured.

Question 10

How should I store the kit?

Accepted Answer

Ship on ice. Store standards QS0/QS1 at −25 to −15 °C and the other components at 2–8 °C. Protect fluorescent reagents and prepared reactions from light.

Question 11

Is this product intended for diagnostic use?

Accepted Answer

No. For research use only. Not for diagnostic or therapeutic procedures.

Parameter	Specification
Biofargo catalog numbers	BF-T1006-S / BF-T1006
Target	RNA
Total target amount per assay	5–100 ng
Initial sample concentration	0.5–100 ng/µL, depending on sample input
Sample input	1–20 µL
Final assay volume	200 µL per tube or well
Working-solution preparation	Prepare fresh working solution by mixing 1 volume of fluorescent reagent with 199 volumes of the supplied buffer (1:200 final dilution).
Incubation	2 minutes at 20–25 °C, protected from light
Signal stability	Up to 3 hours, protected from light
Instrument formats	Biofargo X1 and X3; Qubit fluorometers with the matching RNA assay; fluorescence microplate readers
Plate-reader settings	620 nm excitation / 660 nm emission
Available formats	BF-T1006-S: 100 reactions; BF-T1006: 1,000 reactions
Intended use	Research use only

Component	Designation	BF-T1006-S (100 reactions)	BF-T1006 (1,000 reactions)	Function
RNA HS Buffer QB	QB	20 mL	200 mL	Working-solution buffer
RNA HS Standard QS0	QS0	100 µL	1 mL	Blank standard; 0 ng/µL
RNA HS Standard QS1	QS1	100 µL	1 mL (4 × 250 µL)	High standard; 10 ng/µL
RNA HS Fluorescent Reagent QF	QF	100 µL	1 mL	Concentrated detection reagent

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Biofargo RNA HS Assay Kit