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Portelite™ Fluorimetric RNA Quantitation Kit*Optimized for Cytocite™ and Qubit™ Fluorometers*

RNA standard curve generated using StrandBrite™ Green Fluorimetric RNA Quantitation Kit. Fluorescence intensity was quantified using green fluorescence channel; regression model was calculated using linear fit.
RNA standard curve generated using StrandBrite™ Green Fluorimetric RNA Quantitation Kit. Fluorescence intensity was quantified using green fluorescence channel; regression model was calculated using linear fit.
Ordering information
Price ()
Catalog Number17658
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Excitation (nm)509
Emission (nm)529
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501

OverviewpdfSDSpdfProtocol


Excitation (nm)
509
Emission (nm)
529
Detecting and quantitating small amounts of RNA is extremely important for a wide variety of biological applications such as measuring yields of in vitro transcribed RNA and measuring RNA concentrations before performing Northern blot analysis, S1 nuclease assays, RNase protection assays, cDNA library preparation, reverse transcription PCR, and differential display PCR. The most commonly used technique for measuring nucleic acid concentration is the determination of absorbance at 260 nm. The absorbance-based method is limited by the interferences caused from proteins, free nucleotides and other UV absorbing compounds. The use of sensitive and selective fluorescent nucleic acid stains alleviates this interference problem. StrandBrite™ RNA quantifying reagent is an ultrasensitive fluorescent nucleic acid stain for quantitating RNA in solution. StrandBrite™ RNA quantifying reagent can detect as little as 5 ng/mL RNA with a CytoCite™ or Qubit™ fluorometer. Our StrandBrite™ Green Fluorimetric RNA Quantitation Kit includes our StrandBrite™ Green nucleic acid stain with an optimized and robust protocol. It provides a convenient method for quantifying RNA in solutions.

Platform


Qubit Fluorometer

Excitation480 nm
Emission530 nm
Instrument specification(s)0.2 mL PCR vial

CytoCite Fluorometer

Excitation480 nm
Emission530 nm
Instrument specification(s)0.2 mL PCR vial

Components


Component A: StrandBrite™ Green0.25 mL (200X in DMSO)
Component B: Assay Buffer1 bottle (50 mL)
Component C: RNA Standard #11 mL (RNA: 0 ng/µL)
Component D: RNA Standard #24 X 250 µL (RNA: 10 ng/µL)

Example protocol


AT A GLANCE

Protocol summary
  1. Prepare StrandBrite™ RNA working solution
  2. Add 190 µL StrandBrite™ RNA working solution into each 0.2 mL PCR tube
  3. Add 10 µL RNA Standards or test samples into each tube
  4. Incubate at room temperature for 2 minutes
  5. Monitor fluorescence with CytoCite™ or Qubit™ fluorometer 

Important
Bring all the kit components at room temperature before starting the experiment.

PREPARATION OF WORKING SOLUTION

StrandBrite™ RNA working solution
Make a 200-fold dilution of StrandBrite™ Green (Component A) in Assay Buffer (Component B). For example, to prepare enough working solution for 8 samples, add 5 µL of StrandBrite™ Green (Component A) into 1 mL of Assay Buffer (Component B).
Note     Protect the working solution from light by covering it with foil or placing it in the dark. We recommend preparing this solution in a plastic container rather than glass, as the dye may adsorb to glass surfaces.  For best results, this solution should be used within a few hours of its preparation.

SAMPLE EXPERIMENTAL PROTOCOL

The acceptable sample volume could be a range from 1~20 µL depending on the estimate concentration of RNA sample. The recommend sample volume is 10 µL with the RNA concentration in 0.01~10 ng/µL range. If other sample volume is being used, please adjust the dilution factor in the concentration calculations.
The following protocol is generated based on10 µL sample volume with the RNA concentration in 0.01~10 ng/µL range.
  1. Add 190 µL StrandBrite™ Green working solution into each CytoCite™ sample tube (#CCT100) or equivalent 0.2 mL PCR tube.
    Note     Use thin-wall, polypropylene, clear 0.2 mL PCR tubes such as#CCT100.
  2. Add 10 µL RNA standard #1 and #2 or test samples into each tube, and then mix by vortexing 2~3 seconds.
  3. Incubate all tubes at room temperature for 2 minutes.
  4. Insert the samples into CytoCite™ or Quibit™ and monitor the fluorescence with green fluorescence channel. Follow the procedure appropriate for CytoCite™ Fluorometer. For instructions, see: https://devices.aatbio.com/documentation/user-manual-for-cytocite-fluorometer 

PREPARATION OF STANDARD CALIBRATION CURVE (Optional)
For StrandBrite™ assays, you have the choice to make a calibration curve with the RNA standards. Here is a brief protocol to generate a customized RNA standard curve:
  1. Perform 1/3 serial dilution with Assay Buffer to get 10, 3, 1, 0.3, 0.1, 0.03, 0.01 and 0 ng/µL RNA standard dilutions using RNA Standard #2.
  2. Add 190 µL StrandBrite™ Green working solution into each tube.
  3. Add 10 µL RNA standards or test samples into each tube, and then mix by vortexing 2~3 seconds.
  4. Incubate the reaction at room temperature for 2 minutes.
  5. Insert the samples into CytoCite™ and monitor the fluorescence with green fluorescence channel. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)509
Emission (nm)529

References


View all 50 references: Citation Explorer
Oncologic Implications of Chronic Hepatitis C Virus Infection.
Authors: Hwang, Jessica P and LoConte, Noelle K and Rice, John P and Foxhall, Lewis E and Sturgis, Erich M and Merrill, Janette K and Torres, Harrys A and Bailey, Howard H
Journal: Journal of oncology practice (2019): 629-637
Outcomes and costs of single-step hepatitis C testing in primary care, Birmingham, United Kingdom.
Authors: Munang, M and Smit, E and Barnett, T and Atherton, C and Tahir, M and Atabani, S F
Journal: Public health (2019): 40-44
Evaluation of the Aptima HCV Quant Dx Assay Using Serum and Dried Blood Spots.
Authors: Weber, Jenna and Sahoo, Malaya K and Taylor, Nathaniel and Shi, Run-Zhang and Pinsky, Benjamin A
Journal: Journal of clinical microbiology (2019)
PARP-1/2 Inhibitor Olaparib Prevents or Partially Reverts EMT Induced by TGF-β in NMuMG Cells.
Authors: Schacke, Michelle and Kumar, Janani and Colwell, Nicholas and Hermanson, Kole and Folle, Gustavo A and Nechaev, Sergei and Dhasarathy, Archana and Lafon-Hughes, Laura
Journal: International journal of molecular sciences (2019)
Tumor-draining lymph nodes demonstrate a suppressive immunophenotype in patients with non-small cell lung cancer assessed by endobronchial ultrasound-guided transbronchial needle aspiration: A pilot study.
Authors: Murthy, Vivek and Katzman, Daniel P and Tsay, Jun-Chieh J and Bessich, Jamie L and Michaud, Gaetane C and Rafeq, Samaan and Minehart, Janna and Mangalick, Keshav and de Lafaille, M A Curotto and Goparaju, Chandra and Pass, Harvey and Sterman, Daniel H
Journal: Lung cancer (Amsterdam, Netherlands) (2019): 94-99
Evaluation of two RNA extraction methods for human parechovirus detection on pediatric stool specimens.
Authors: Bergallo, Massimiliano and Montanari, Paola and Daprà, Valentina and Cuccu, Roberto and Bonamin, Stefano and Gabiano, Clara and Galliano, Ilaria
Journal: Minerva pediatrica (2018)
Comparison of the Aptima HIV-1 Quant Dx assay with the COBAS AmpliPrep/COBAS TaqMan HIV-1 v2.0 Test for HIV-1 viral load quantification in plasma samples from HIV-1-infected patients.
Authors: Longo, Serena and Bon, Isabella and Musumeci, Giuseppina and Bertoldi, Alessia and D'Urbano, Vanessa and Calza, Leonardo and Re, Maria Carla
Journal: Health science reports (2018): e31
A pilot study on primary cultures of human respiratory tract epithelial cells to predict patients' responses to H7N9 infection.
Authors: Huang, Chung-Guei and Lee, Li-Ang and Wu, Yi-Cheng and Hsiao, Mei-Jen and Horng, Jim-Tong and Kuo, Rei-Lin and Huang, Chih-Heng and Lin, Ya-Chu and Tsao, Kuo-Chien and Chen, Min-Chi and Chen, Tse-Ching and Shih, Shin-Ru
Journal: Oncotarget (2018): 14492-14508
Evaluation of dried blood spot as an alternative sample collection method for hepatitis C virus RNA quantitation and genotyping using a commercial system.
Authors: Mahajan, Supriya and Choudhary, Manish Chandra and Kumar, Guresh and Gupta, Ekta
Journal: Virusdisease (2018): 141-146
c9orf72 Disease-Related Foci Are Each Composed of One Mutant Expanded Repeat RNA.
Authors: Liu, Jing and Hu, Jiaxin and Ludlow, Andrew T and Pham, Jacqueline T and Shay, Jerry W and Rothstein, Jeffrey D and Corey, David R
Journal: Cell chemical biology (2017): 141-148