StrandBrite™ Green Fluorimetric RNA Quantitation Kit *Optimized for Microplate Readers*


Tests: 1000
Price:
$732

Overview 

Emission (nm): 545     

Excitation (nm): 490

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 (nm): 490

Emission (nm): 545 

Cutoff (nm): 515  

Recommended Plate: Solid Black

 

Components

Component A: StrandBrite™ Green: 1 vial (0.5 mL, 200X in DMSO)

Component B: 10X Assay Buffer: 1 bottle (20 mL)

Component C: Ribosomal RNA  Standard: 1 vial (0.2mL, 100 µg/mL)

 

Example protocol

At a glance

Protocol summary

  1. Add 100 µL RNA standards or test samples
  2. Add 100 µL StrandBrite Green™ working solution
  3. Incubate at RT for 2-5 minutes
  4. Monitor the fluorescence at Ex/Em=490/545 nm

Important notes
The following protocol is an example for quantifying RNA with StrandBrite™ Green. Allow all the components to warm to room temperature before opening. Prevent RNase contamination of the StrandBrite™ reagent and kit components. Always use clean disposable gloves while handling all materials. Use nuclease-free water and sterile, disposable polypropylene plastic ware for reagent preparation. Caution: No data are available for addressing the mutagenicity or toxicity of StrandBrite™ Green RNA stain. Because this reagent binds to nucleic acids, it should be treated as a potential mutagen and handled with appropriate care. The DMSO stock solution should be handled with caution as DMSO is known to facilitate the entry of organic molecules into tissues.

Preparation of stock solution

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.

1. 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

Add 10 µL of 100 µg/mL Ribosomal 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.

References

 

 

Inhibitors of Streptococcus pneumoniae surface endonuclease EndA discovered by high-throughput screening using a PicoGreen fluorescence assay
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
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
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
Authors: Abiodun OO, Gbotosho GO, Ajaiyeoba EO, Happi CT, Hofer S, Wittlin S, Sowunmi A, Brun R, Oduola AM.
Journal: Parasitol Res (2010): 933

Metal-enhanced PicoGreen fluorescence: application to fast and ultra-sensitive pg/ml DNA quantitation
Authors: Dragan AI, Bishop ES, Casas-Finet JR, Strouse RJ, Schenerman MA, Geddes CD.
Journal: J Immunol Methods (2010): 95

Quantification of dsDNA using the Hitachi F-7000 Fluorescence Spectrophotometer and PicoGreen dye
Authors: Moreno LA, Cox KL.
Journal: J Vis Exp. (2010)

Development and characterization of a novel host cell DNA assay using ultra-sensitive fluorescent nucleic acid stain "PicoGreen"
Authors: Ikeda Y, Iwakiri S, Yoshimori T.
Journal: J Pharm Biomed Anal (2009): 997

Enhanced DNA dynamics due to cationic reagents, topological states of dsDNA and high mobility group box 1 as probed by PicoGreen
Authors: Noothi SK, Kombrabail M, Kundu TK, Krishnamoorthy G, Rao BJ.
Journal: FEBS J (2009): 541

Factors affecting quantification of total DNA by UV spectroscopy and PicoGreen fluorescence
Authors: Holden MJ, Haynes RJ, Rabb SA, Satija N, Yang K, Blasic JR, Jr.
Journal: J Agric Food Chem (2009): 7221

Label-free DNA sequence detection with enhanced sensitivity and selectivity using cationic conjugated polymers and PicoGreen
Authors: Ren X, Xu QH.
Journal: Langmuir (2009): 43

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