StrandBrite™ Green Fluorimetric RNA Quantitation Kit *Optimized for Microplate Readers*
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Spectral properties
Excitation (nm) | 509 |
Emission (nm) | 527 |
Storage, safety and handling
H-phrase | H303, H313, H340 |
Hazard symbol | T |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R68 |
UNSPSC | 41116134 |
Alternative formats
StrandBrite™ Green Fluorimetric RNA Quantitation Kit |
StrandBrite™ Green Fluorimetric RNA Quantitation Kit *High Selectivity* |
Related products
StrandBrite™ Green RNA Quantifying Reagent *200X DMSO Solution* |
Overview | ![]() ![]() |
See also: Gene Expression Analysis & Genotyping, Genome Editing & CRISPR, Coronavirus (SARS-CoV-2), RNA Immunoprecipitation (RIP), RNA Purification & Analysis
Excitation (nm) 509 | Emission (nm) 527 |
Detecting and quantitating small amounts of RNA is extremely important for a wide variety of molecular biology procedures 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 major disadvantage of the absorbance-based method is the interferences caused by proteins, free nucleotides and other UV absorbing compounds. The use of sensitive, 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 fluorescence microplate reader or 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
Fluorescence microplate reader
Excitation | 490 nm |
Emission | 545 nm |
Cutoff | 515 nm |
Recommended plate | Solid black |
Components
Example protocol
AT A GLANCE
Protocol Summary
- Add 100 µL RNA standards or test samples
- Add 100 µL StrandBrite Green™ working solution
- Incubate at RT for 2-5 minutes
- Monitor the fluorescence at Ex/Em=490/545 nm
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Assay Buffer stock solution (1X)
Dilute the concentrated 10X Assay Buffer (Component B) to 1X Assay Buffer stock solution with sterile, distilled, nuclease-free water.PREPARATION OF STANDARD SOLUTION
For convenience, use the Serial Dilution Planner:
https://www.aatbio.com/tools/serial-dilution/17655
https://www.aatbio.com/tools/serial-dilution/17655
RNA standard
Add 10 µL of 100 µg/mL RNA Standard (Component C) to 990 µL of 1X Assay Buffer to have 1 µg/mL RNA standard solution (RS7). Then perform 1:3 serial dilutions to get remainder serial dilutions (RS1 - RS6).PREPARATION OF WORKING SOLUTION
Prepare StrandBrite™ Green working solution by making a 200-fold dilution of the concentrated DMSO solution in 1X Assay Buffer. For example, add 50 μL of StrandBrite™ Green (Component A) into 10 mL of 1X Assay Buffer. Protect the working solution from light by covering it with foil or placing it in the dark.
Note We recommend preparing this solution in a plastic container rather than glass, as the dye may adsorb to glass surfaces. For best results, use promptly, within a few hours of its preparation.
Note We recommend preparing this solution in a plastic container rather than glass, as the dye may adsorb to glass surfaces. For best results, use promptly, within a few hours of its preparation.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of RNA standards and test samples in a solid black 96-well microplate. SD = RNA standard (SD1 - SD7, 1 to 1000 ng/mL); BL = blank control; TS = test sample.
Table 2. Reagent composition for each well.
BL | BL | TS | TS |
SD1 | SD1 | ... | ... |
SD2 | SD2 | ... | ... |
SD3 | SD3 | ||
SD4 | SD4 | ||
SD5 | SD5 | ||
SD6 | SD6 | ||
SD7 | SD7 |
Well | Volume | Reagent |
SD1 - SD7 | 100 µL | serial dilution (1 to 1000 ng/mL) |
BL | 100 µL | TE Buffer |
TS | 100 µL | sample |
- Prepare RNA standards (RS), blank controls (BL), and test samples (TS) in a 96-well solid black microplate according to the layout provided in Table 1 and Table 2. For a 384-well plate, add 25 µL of reagent per well instead of 100 µL.
- Add 100 µL of StrandBrite™ Green working solution to each well of the RNA standard, blank control, and test samples to make the total RNA assay volume of 200 µL/well. For a 384-well plate, add 25 µL of working solution into each well instead, for a total volume of 50 µL/well.
- Incubate the reaction at room temperature for 2 to 5 minutes; protect from light.
- Monitor the fluorescence increase with a spectrofluorometer at Ex/Em = 490/545 nm (cutoff at 515 nm).
Note To minimize photobleaching, keep the time for fluorescence measurement constant across all samples.
Images
Citations
View all 1 citations: Citation Explorer
Anticancer Activity of Reconstituted Ribonuclease S-Decorated Artificial Viral Capsid
Authors: Liang, Yingbing and Furukawa, Hiroto and Sakamoto, Kentarou and Inaba, Hiroshi and Matsuura, Kazunori
Journal: ChemBioChem (2022): e202200220
Authors: Liang, Yingbing and Furukawa, Hiroto and Sakamoto, Kentarou and Inaba, Hiroshi and Matsuura, Kazunori
Journal: ChemBioChem (2022): e202200220
References
View all 31 references: Citation Explorer
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Journal: J Biomol Screen (2013): 247
Authors: Peterson EJ, Kireev D, Moon AF, Midon M, Janzen WP, Pingoud A, Pedersen LC, Singleton SF.
Journal: J Biomol Screen (2013): 247
Validation of a PicoGreen-based DNA quantification integrated in an RNA extraction method for two-dimensional and three-dimensional cell cultures
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Authors: Chen Y, Sonnaert M, Roberts SJ, Luyten FP, Schrooten J.
Journal: Tissue Eng Part C Methods (2012): 444
Characterization of PicoGreen interaction with dsDNA and the origin of its fluorescence enhancement upon binding
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Authors: Dragan AI, Casas-Finet JR, Bishop ES, Strouse RJ, Schenerman MA, Geddes CD.
Journal: Biophys J (2010): 3010
Comparison of SYBR Green I-, PicoGreen-, and [3H]-hypoxanthine-based assays for in vitro antimalarial screening of plants from Nigerian ethnomedicine
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Journal: Parasitol Res (2010): 933
Metal-enhanced PicoGreen fluorescence: application to fast and ultra-sensitive pg/ml DNA quantitation
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Journal: J Immunol Methods (2010): 95
Authors: Dragan AI, Bishop ES, Casas-Finet JR, Strouse RJ, Schenerman MA, Geddes CD.
Journal: J Immunol Methods (2010): 95
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Journal: J Vis Exp. (2010)
Authors: Moreno LA, Cox KL.
Journal: J Vis Exp. (2010)
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Authors: Holden MJ, Haynes RJ, Rabb SA, Satija N, Yang K, Blasic JR, Jr.
Journal: J Agric Food Chem (2009): 7221
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Authors: Ikeda Y, Iwakiri S, Yoshimori T.
Journal: J Pharm Biomed Anal (2009): 997
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Authors: Noothi SK, Kombrabail M, Kundu TK, Krishnamoorthy G, Rao BJ.
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Authors: Ren X, Xu QH.
Journal: Langmuir (2009): 43
Application notes
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