Qubit® Fluorometer Technical Resources
Here you can find the most up-to-date technical resources for your Qubit® 3.0 Fluorometer, including a variety of application and technical notes such as Qubit® Quantitation Platform vs. the Nanodrop® Spectrophotometer and Testing for Bias in the Qubit® RNA Assay, and the Qubit® Frequently Asked Questions (FAQs).
Technical and application notes
- Comparison of fluorescence-based quantitation with UV absorbance measurements—Qubit® fluorometric quantitation vs. spectrophotometer measurements
- Qubit® dsDNA assay specificity in the presence of single-stranded DNA
- Comparison of Quant-iT™ and Qubit® DNA quantification assays for accuracy and precision
- The use of glycogen and GlycoBlue™ reagent in Qubit® DNA and RNA assays as measured on the Qubit® Fluorometer
- Qubit® Fluorometer vs. Quantus™ Fluorometer
- Accurate and sensitive protein quantitation—Comparison of the Qubit® Protein Assay for the Qubit® Fluorometer and other conventional protein assays
- Testing for bias in the Qubit™ RNA Assay using single- and double-stranded RNA homopolymers
From the Qubit® 3.0 Fluorometer Home screen, touch Settings, then touch About instrument to verify the version of software currently on your instrument. If your instrument requires a software upgrade, you can transfer the latest software onto the Qubit® 3.0 Fluorometer using a USB drive or USB cable and install it by touching Settings, and then Update software. For more detailed instructions, please consult the Qubit® 3.0 product manual.
- Download the latest version (v1.02) of the Qubit® 3.0 instrument software
IMPORTANT: Right (or control) click on the link above and choose "Save file as." The file extension of the saved file should be .pak.
Connecting the Qubit® 3.0 to a computer running Windows® XP operating system
Connecting the Qubit® 3.0 Fluorometer to a computer running Windows® XP operating system requires installation of a driver. You can download both the driver and installation instructions below.
Below are some frequently asked questions and answers regarding the Qubit® Fluorometer and Qubit® Fluorometric Quantitation. If you have a question that is not listed below or need additional information, please contact our Technical Support department.
Q. I already have a Nanodrop® instrument. Why should I use the Qubit® Fluorometer?
A. The Nanodrop® instrument uses UV absorbance, which cannot distinguish between DNA, RNA, free nucleotides, and other contaminants.The Qubit® assays are more sensitive than the NanoDrop® instrument. NanoDrop® Spectrophotometer full-spectrum absorption readings can give peaks revealing the presence of contaminants. This may be useful information for downstream applications in which the contaminants might be detrimental. When used together, the Qubit® Fluorometer can accurately measure low concentrations of sample and the Nanodrop spectrophotometer can measure contaminants.
Q. Why are my UV absorbance readings higher than the Qubit® Fluorometer readings?
A. UV absorbance readings measure anything that absorbs at 260 nm, including DNA, RNA, protein, free nucleotides, and excess salt. Qubit® Fluorometric Quantitation only measures the molecule you are interested in, so the number is almost always lower than the A260 reading.
Q. Can the Qubit® Fluorometer give an indication of sample quality?
A. Yes. You can use one of the DNA quantitation kits to measure DNA concentration, one of the RNA kits to measure RNA concentration, and the protein kit to measure protein concentration. Use a combination of whichever kits you need for the biomolecule you are interested in and the contaminant of concern. Together, they give you accurate information about how much DNA, RNA, and protein you have in your sample.
Q. I already have a Qubit® Fluorometer, is Qubit® 3.0 Fluorometer any different?
A. The Qubit® 3.0 Fluorometer employs a large, robust color touch screen for seamless workflow navigation and exports data to a USB drive or directly to your computer via a USB cable for efficient data management. Also, the instrument can be personalized to show only the frequently used assays, to add new assays, including user-defined assays created with the MyQubit assay design tool, and to display in the language of your choice including English, French, Spanish, Italian, German, simplified Chinese, and Japanese.
Q. Can I use my old Quant-iT™ Kits labeled “for use with Qubit® Fluorometer” with the Qubit® 3.0 Fluorometer?
A. Yes, these kits will work with all Qubit® Fluorometers.
Q. How many lines of data can the Qubit® Fluorometers store?
A. The Qubit® 3.0 Fluorometer can store up to 1,000 samples’ worth of data in a .csv file.
The Qubit® 2.0 Fluorometer can store up to 200 lines of data in a .csv file.
The original Qubit® Fluorometer can store up to 20 lines of data in a .csv file.
Q. Do I have to use new standards every time?
A. No. But we do recommend using new standards every time you make a new working solution, so that the working solution used in your standards is the same as that used in your samples.
Q. How long can the diluted standards be saved and reused? What if there is evaporation over time (obvious or not obvious)?
A. The diluted standards can be used for up to three hours if using the same working solution for the samples.
Q. Can Qubit® Fluorometric Quantitation quantify plasmids?
A. Yes. Use the Qubit® DNA BR assay for a typical plasmid miniprep with lots of DNA (over 50 ng/μL). Use the Qubit® DNA HS assay for “plasmid rescue” or methods that yield only small amounts of DNA.
Q. Is there a difference in signal between supercoiled and relaxed DNA?
A. Yes. For supercoiled DNA, we recommend nicking the DNA so it is not supercoiled, or using supercoiled DNA as Standard 2.
Q. Does the Qubit® Protein Assay work well in the presence of detergents?
A. It is compatible with very small amounts of detergent. See “Contaminants Tolerated by the Qubit® Protein Assay,” Table 2 on page 6 of the Qubit® Protein Assay Kit product manual for specific amounts.
Q. How long does the lamp last? How do I change the lamp?
A. There are two light sources in the Qubit® 2.0 and 3.0 Fluorometers—both are LEDs. They are expected to last at least 5 years.
Q. Can I repair my Qubit® Fluorometer?
A. No. The warranty will be voided if the instrument is disassembled or a customer has attempted to repair the instrument.
Q. What if my Qubit® Fluorometer fails under warranty?
A. We will replace your Qubit® Fluorometer. Please contact Technical Support for details.
Q. What kind of tubes do I need to buy?
A. Use thin-wall, clear 0.5 mL PCR tubes such as Qubit® assay tubes or Axygen PCR-05-C tubes (VWR, part number 10011-830).
- Adams RI, Miletto M, Lindow SE, Taylor JW, Bruns TD (2014) Airborne Bacterial Communities in Residences: Similarities and Differences with Fungi. PLoS ONE 9(3):e91283 doi:10.1371.
- Callaway JL, Huang S, Karampetsou E, Crolla JA (2014) Perspective on the technical challenges involved in the implementation of array-CGH in prenatal diagnostic testing. Mol Biotechnol 56(4):312–8.
- de Bourcy CFA, De Vlaminck I, Kanbar JN, Wang J, Gawad C, et al. (2014) A Quantitative Comparison of Single-Cell Whole Genome Amplification Methods. PLoS ONE 9(8):e105585. doi:10.1371.
- Hewitson L, Thissen JB, Gardner SN, McLoughlin KS, Glausser MK, Jaing CJ, (2014) Screening of Viral Pathogens from Pediatric Ileal Tissue Samples after Vaccination. Advances in Virology, vol. 2014, Article ID 720585 10 pp doi:10.1155.
- Kulkarni AD, Kiron V, Rombout JH et al. (2014) Protein profiling in the gut of Penaeus monodon gavaged with oral WSSV-vaccines and live white spot syndrome virus. Proteomics 14: 1660–1673.
- Sugiyama A, Ueda Y, Zushi T, Takase H, Yazaki K (2014) Changes in the Bacterial Community of Soybean Rhizospheres during Growth in the Field. PLoS ONE 9(6):e100709. doi:10.1371.
- Deben C, Zwaenepoel K, Boeckx C et al. (2013) Expression Analysis on Archival Material Revisited: Isolation and Quantification of RNA Extracted From FFPE Samples. Diagn Mol Pathol 22(1):59–64.
- Fitzsimons MS, Novotny M, Lo CC et al. (2013) Nearly finished genomes produced using gel microdroplet culturing reveal substantial intraspecies genomic diversity within the human microbiome. Genome Res 23(5):878–88.
- Hebling CM, McFarland MA, Callahan JH, Ross MM (2013) Global Proteomic Screening of Protein Allergens and Advanced Glycation Endproducts in Thermally Processed Peanuts. J Agric Food Chem 61(24):5638–5648.
- Bundo M, Sunaga F, Ueda J et al. (2012) A systematic evaluation of whole genome amplification of bisulfite-modified DNA. Clin Epigenetics doi:10.1186.
- Gibson WT, Hood RL, Zhan SH et al. (2012) Mutations in EZH2 Cause Weaver Syndrome. Am J Hum Genet 90(1):110–118.
- Kondragunta B, Joshi BH, Han J et al. (2012) Bioreactor environment-sensitive sentinel genes as novel metrics for cell culture scale-down comparability. Biotechnol Prog 28(5):1138–1151.
- Sharp CE, Stott MB, Dunfield PF (2012) Detection of autotrophic verrucomicrobial methanotrophs in a geothermal environment using stable isotope probing. Front Microbiol doi:10.3389.
- Valenzuela J, Mazurie A, Carlson RP et al. (2012) Potential role of multiple carbon fixation pathways during lipid accumulation in Phaeodactylum tricornutum. Biotechnol Biofuels 5(1):40.
- Yang WC, Pate KG, Wong HE, Swartz JR (2012) Simplifying and streamlining Escherichia coli-based cell-free protein synthesis. Biotechnol Prog 28(2):413–420.
- Binder M, Eberle F, Seitz S et al. (2011) Molecular Mechanism of Signal Perception and Integration by the Innate Immune Sensor Retinoic Acid-inducible Gene-I (RIG-I). J Biol Chem 286(31):27278–27287.
- Debreuil J, Tambutté S, Zoccola D et al. (2011) Comparative analysis of the soluble organic matrix of axial skeleton and sclerites of Corallium rubrum: Insights for biomineralization. Comp Biochem Physiol B Biochem Mol Biol. Part B 159(1):40–48.
- Ferreira JA, Daniel-da-Silva AL, Alves RM et al. (2011) Synthesis and Optimization of Lectin Functionalized Nanoprobes for the Selective Recovery of Glycoproteins from Human Body Fluids. Anal Chem 83(18):7035–7043.
- Hares K, Kemp K, Gray E et al. (2011) Neurofilament dot blot assays: Novel means of assessing axon viability in culture. J Neurosci Methods 198(2):195–203.
- Holloway, CM, McIntyre CK (2011) Post-training disruption of Arc protein expression in the anterior cingulate cortex impairs long-term memory for inhibitory avoidance training. Neurobiol Learn and Mem 95:425–432.
- Kwezi L, Ruzvidzo O, Wheeler JI et al. (2011) The Phytosulfokine (PSK) Receptor Is Capable of Guanylate Cyclase Activity and Enabling Cyclic GMP-dependent Signaling in Plants. J Biol Chem 286(25):22580–22588.
- Pérez NG, Nolly MB, Roldan MC et al. (2011) Silencing of NHE-1 blunts the slow force response to myocardial stretch. J Appl Physiol 111(3):874–880.
- Recouvreux MV, Guida MC, Rifkin DB et al. (2011) Active and Total Transforming Growth Factor-β1 Are Differentially Regulated by Dopamine and Estradiol in the Pituitary. Endocrinology 152(7):2722–2730.
- Svedružić D, Blackburn JL, Tenent RC et al. (2011) High-Performance Hydrogen Production and Oxidation Electrodes with Hydrogenase Supported on Metallic Single-Wall CarbonNanotube Networks. J Am Chem Sod 133(12):4299–4306.
- Couto GH et al. (2010) Isolation of a novel lipase from a metagenomic library derived from mangrove sediment from the south Brazilian coast. Genetic Mol Res 9(1):514–523.
- Darnell M et al. (2010) Investigation of the involvement of p-glycoprotein and multidrug resistance-associated protein 2 in the efflux of ximelagatran and its metabolites by using short hairpin RNA knockdown Caco-2 cells. Drug Metab Disp 38:491–497.
- Dunn AK et al. (2010) The alternative oxidase (AOX) gene in Vibrio fischeri is controlled aby NsrR and upregulated in response to nitric oxide. Molecular Microbiol 77(1):44–55.
- Fajardo-Cavazos P et al. (2010) Exposure of DNA and Bacillus subtilis Spores to Simulated Martian Environments: Use of Quantitative PCR (qPCR) to Measure Inactivation Rates of DNA to Function as a Template Model. Astrobiology 10(4):403–411.
- Filigheddu N, et al. (2010) Differential Expression of MicroRNAs between Eutopic and Ectopic Endometrium in Ovarian Endometriosis. J Biomed Biotech 2010:369549.
- Gupta RK, Srivastava BS, Srivastava R (2010) Comparative expression analysis of rpf-like genes of Mycobacterium tuberculosis H37Rv under different physiological stress and growth. conditions Microbiology 156:2714–2722.
- Lee, SE et al. (2010) Prevalence of Toxoplasmosis gondii infection in stray and household cats in regions of Seoul, Korea. Korean J Parasitol 48(3):267–270.
- Podolyan, A., et.al. (2010) Identification of the lipoxygenase gene family from Vitis vinifera and biochemical characterization of two 13-lipoxygenases expressed in grape berries of Sauvignon Blanc. Functional Plant Biol 37:767–784.
- Regueiro BJ et.al. (2010) Automated extraction improves multiplex molecular detection of infection in septic patients. PLoS One 5(10):e13387.
- Simmons M, Donovan DM, Siragusa GR, Seal BR (2010) Recombinant Expression of Two Bacteriophage Proteins That Lyse Clostridium perfringens and Share Identical Sequences in the C-Terminal Cell Wall Binding Domain of the Molecules but Are Dissimilar in Their N-Terminal Active Domains. J Agric Food Chem 58(19):10330–10337.
- Tennico YH et.al. (2010) On-chip aptamer-based sandwich assay for thrombin detection employing magnetic beads and quantum dots. Anal Chem 82:5591–5597.
Qualifying DNA preparations for next-generation sequencing (NGS)
Prior to committing the effort and resources required for next-generation sequencing (NGS), it’s important for researchers to be able to quantify and assess the quality of the DNA starting material. In a 2013 publication, Simbolo et al. reported a reliable and cost-effective method to qualify various DNA preparations for NGS, including those derived from frozen tissue and FFPE samples. Of the instruments they tested, their data showed that DNA quantification results obtained using the Qubit® Fluorometer were highly reproducible and were consistent with qPCR data for DNA quantity—even for partially degraded DNA from FFPE samples.
For Research Use Only. Not for use in diagnostic procedures.