Qubit fluorimeter

Manufactured by Qiagen

The Qubit fluorimeter is a compact and easy-to-use instrument designed for quantifying nucleic acids and proteins. It utilizes fluorescent dyes to accurately measure the concentration of samples, providing reliable results with a simple workflow.

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Lab products found in correlation

Tetro reverse transcription kit, by Meridian Bioscience (3 mentions) Rna br kit, by Qiagen (3 mentions) Oligo dt 15 mers, by Integrated DNA Technologies (3 mentions) Sybr green mix, by Agilent Technologies (3 mentions) Tissuelyser 2, by Qiagen (3 mentions) Qiazol reagent, by Qiagen (3 mentions) Rnalater, by Thermo Fisher Scientific (3 mentions) Lightcycler 480 2, by Roche (3 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions)

Spelling variants of this product

Qubit 2.0 Fluorimeter (2 citations)

4 protocols using qubit fluorimeter

DNA Extraction and Quantification

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Timing: 2–4 h

DNA samples are extracted with Qiagen DNeasy blood and tissue kit (Cat. No. 69504) or any other high-quality DNA extraction kit according to the manufacturer's protocol.

Follow the protocol of Qiagen DNeasy blood and tissue kit.

Measure the DNA concentration of the samples using Qubit fluorimeter and perform the necessary dilutions to obtain a minimum final concentration of 15 ng/μL.

Pause point: isolated DNA can be stored at −20°C for use in subsequent steps.

Note: Run the DNA samples on 1% agarose gel electrophoresis to verify the integrity of the extracted DNA.

CRITICAL: It is recommended to use the Qubit™ Fluorometric DNA Quantification system (Thermo Fisher Scientific) to measure the DNA concentration, and nanodrop to check purity (a ratio of absorbance at 260 nm vs 280 nm of ∼1.8 is considered optimal). To prepare the Qubit™ reagents, follow the instruction of its kit. It is recommended to consider the results of Qubit™ measurements as the basis for calculating the amount of DNA. The concentrations measured should allow the addition of 400 ng of each individual DNA to be digested with the PstI enzyme within a reaction volume of 30 μl (see below of the detailed components of the PstI digestion reaction).

Rezaei S., Uffenorde J., Gimm O., Hosseinpour Feizi M.A., Miemczyk S., Coutinho L.L., Jensen P., Guerrero-Bosagna C, & Pértille F. (2022). GBS-MeDIP: A protocol for parallel identification of genetic and epigenetic variation in the same reduced fraction of genomes across individuals. STAR Protocols, 3(1), 101202.

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Quantitative RT-PCR Analysis of Gene Expression

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Tissue samples stored in RNAlater (Thermo Fisher Scientific) were processed for RNA extraction using a TissueLyser II and QIAzol reagent (QIAGEN). Isolated RNA was quantified using a Qubit fluorimeter and RNA BR kit (QIAGEN). cDNA was synthesized using Tetro reverse transcription kit (Bioline) and oligo dT 15-mers (Integrated DNA Technologies). Quantitative real-time PCR was performed using SYBR™ green mix (Agilent Technologies) and a LightCycler 480 II (Roche). A list of primer sequences used are shown in Table 2. Gene expression levels were determined by second derivative maxima using standard curves (LightCycler software) and expressed relative to the housekeeping gene Rpl13a.

Chenery A.L., Rosini S., Parkinson J.E., Ajendra J., Herrera J.A., Lawless C., Chan B.H., Loke P., MacDonald A.S., Kadler K.E., Sutherland T.E, & Allen J.E. (2021). IL-13 deficiency exacerbates lung damage and impairs epithelial-derived type 2 molecules during nematode infection. Life Science Alliance, 4(8), e202001000.

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Comprehensive RNA extraction and quantitative PCR

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Tissue samples stored in RNAlater (Thermo Fisher Scientific) were processed for RNA extraction using a TissueLyser II and QIAzol reagent (Qiagen). Isolated RNA was quantified using a Qubit fluorimeter and RNA BR kit (Qiagen). cDNA was synthesized using Tetro reverse transcription kit (Bioline) and oligo dT 15-mers (Integrated DNA Technologies). Quantitative real-time PCR was performed using SYBR Green mix (Agilent Technologies) and a LightCycler 480 II (Roche). A list of primer sequences used is shown in Table II.

Chenery A.L., Alhallaf R., Agha Z., Ajendra J., Parkinson J.E., Cooper M.M., Chan B.H., Eichenberger R.M., Dent L.A., Robertson A.A., Kupz A., Brough D., Loukas A., Sutherland T.E., Allen J.E, & Giacomin P.R. (2019). Inflammasome-Independent Role for NLRP3 in Controlling Innate Antihelminth Immunity and Tissue Repair in the Lung. The Journal of Immunology Author Choice, 203(10), 2724-2734.

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Quantitative RNA Analysis from Tissue Samples

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Tissue samples stored in RNAlater (Thermo Fisher Scientific) were processed for RNA extraction using a TissueLyser II and QIAzol reagent (Qiagen). Isolated RNA was quantified using a Qubit fluorimeter and RNA BR kit (Qiagen). cDNA was synthesized using Tetro reverse transcription kit (Bioline) and oligo dT 15-mers (Integrated DNA Technologies). Quantitative real-time PCR was performed using SYBR green mix (Agilent Technologies) and a LightCycler 480 II (Roche). A list of primer sequences used are shown in Table II.

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