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Portelite™ Fluorimetric Total Nucleic Acid Quantitation Kit *Optimized for Cytocite™ and Qubit™ Fluorometers*

Comparison of total nucleic acid dose response using the Qubit™ fluorometer (blue) or CytoCite™ fluorometer (red).
Comparison of total nucleic acid dose response using the Qubit™ fluorometer (blue) or CytoCite™ fluorometer (red).
Ordering information
Price ()
Catalog Number17631
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
Portelite™ Fluorimetric Total Nucleic Acid Quantitation Kit is designed to rapidly measure the total amounts of nucleic acids, including double-stranded DNA (dsDNA), single-stranded DNA (ssDNA) and RNA in a sample. The kit has all the essential reagents, including Helixyte™ Green ssDNA reagent, dilution buffer, and prediluted DNA standards. Helixyte™ Green All reagent is a sensitive fluorescent nucleic acid probe for measuring the total amounts of nucleic acids in a sample that may contain double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), RNA and long oligonucleotides. Helixyte™ Green All reagent indiscriminately binds to dsDNA, ssDNA and RNA. Portelite™ Fluorimetric Total Nucleic Acid Quantitation Kit is optimized for measuring the total amounts of nucleic acids with CytoCite™ or Qubit® fluorometers.

Platform


Qubit Fluorometer

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

CytoCite Fluorometer

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

Components


Component A: Helixyte™ Green All1 vial (0.25 mL-200X DMSO)
Component B: Assay Buffer1 bottle (100 mL)
Component C: Nucleic Acid Standard #11 vial (1 mL, 0 ng/µL)
Component D: Nucleic Acid Standard #21 vial (1 mL, 10 ng/µL)

Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare a Helixyte™ Green All working solution
  2. Add 190 µL of 1X Helixyte™ Green All working solution into each 0.2 mL PCR tube
  3. Add 10 µL of Nucleic Acid Standards or test samples into each tube
  4. Incubate at room temperature for 2 minutes
  5. Monitor the fluorescence intensity with CytoCite™ fluorometer or Qubit™ fluorometer 

Important
All kit components must be brought to room temperature before starting the experiment.

PREPARATION OF WORKING SOLUTION

Helixyte™ Green All working solution
Make a 200-fold dilution of Helixyte™ Green All reagent (Component A) with Assay Buffer (Component B). For example, to prepare enough working solution for 5 samples, add 5 μL of Helixyte™ Green All (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. It’s recommended to prepare the solution in a plastic container rather than a glass container, as the dye may adsorb to the glass surface.  For best results, this solution should be used within a few hours after the dilution.

SAMPLE EXPERIMENTAL PROTOCOL

The acceptable range for the sample volume could be 1~20 μL depending on the estimated concentration of the Nucleic Acid sample.
The following protocol is generated based on a sample volume of 10 μL.
  1. Add 190 µL of 1X Helixyte™ Green All working solution into each Cytocite™ sample tube (#CCT100) or the equivalent 0.2 mL PCR tube.
    Note     Use thin-wall, polypropylene, clear 0.2 mL PCR tubes such as AAT Cat#CCT100.
  2. Add 10 μL of Nucleic Acid Standards or test samples into each tube, and then mix by vortexing for 2~3 seconds.
  3. Incubate all tubes at room temperature for 2 minutes.
  4. Insert the samples into CytoCite™ or Qubit™ and monitor the fluorescence intensity with the green fluorescence channel. Follow the appropriate procedures for CytoCite™ Fluorometer. See the link below for detailed instructions: https://devices.aatbio.com/documentation/user-manual-for-cytocite-fluorometer 

Preparation of Standard Calibration Curve
For Portelite™ assays, you have the choice to make a calibration curve with the Nucleic Acid Standards. Here is a brief protocol to generate a customized DNA standard curve.
  1. Perform a 1:2 serial dilution: Add 10 ng/μL Nucleic Acid Standard #2 (Component D) into the Assay Buffer (Component B) to get 10, 5, 2.5, 1.25, 0.62, 0.31, 0.15 ng/µL DNA standard dilutions.
  2. Add 190 µL of the Helixyte™ Green All working solution into each tube.
  3. Add 10 µL of standards into a 0.2 mL PCR tube and then mix by vortexing for 2∼3 seconds.
  4. Incubate the reaction at room temperature for 2 minutes.
  5. Insert the samples into CytoCite™ and monitor the fluorescence intensity with the green fluorescence channel. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)509
Emission (nm)529

References


View all 13 references: Citation Explorer
Accurate bulk quantitation of droplet digital PCR.
Authors: Sun, Chen and Liu, Leqian and Vasudevan, Harish N and Chang, Kai-Chun and Abate, Adam R
Journal: bioRxiv : the preprint server for biology (2021)
Nucleic Acid Quantitation with Log-Linear Response Hybridization Probe Sets.
Authors: Wu, Lucia R and Fang, John Z and Khodakov, Dmitriy and Zhang, David Yu
Journal: ACS sensors (2020): 1604-1614
Nucleic Acid Extraction from Human Biological Samples.
Authors: Mullegama, Sureni V and Alberti, Michael O and Au, Cora and Li, Yan and Toy, Traci and Tomasian, Vanina and Xian, Rena R
Journal: Methods in molecular biology (Clifton, N.J.) (2019): 359-383
MICROFLUIDIC DEVICES FOR LABEL-FREE AND NON-INSTRUMENTED QUANTITATION OF UNAMPLIFIED NUCLEIC ACIDS BY FLOW DISTANCE MEASUREMENT.
Authors: Chatterjee, Debolina and Mansfield, Danielle S and Woolley, Adam T
Journal: Analytical methods : advancing methods and applications (2014): 8173-8179
Helicase-dependent amplification of nucleic acids.
Authors: Cao, Yun and Kim, Hyun-Jin and Li, Ying and Kong, Huimin and Lemieux, Bertrand
Journal: Current protocols in molecular biology (2013): 15.11.1-15.11.12
Microvolume quantitation of nucleic acids.
Authors: Desjardins, Philippe R and Conklin, Deborah S
Journal: Current protocols in molecular biology (2011): 3J
Concentration determination of nucleic acids and proteins using the micro-volume BioSpec-nano-spectrophotometer.
Authors: Sukumaran, Suja
Journal: Journal of visualized experiments : JoVE (2011)
NanoDrop microvolume quantitation of nucleic acids.
Authors: Desjardins, Philippe and Conklin, Deborah
Journal: Journal of visualized experiments : JoVE (2010)
Comparison of two real-time PCR methods for detection of ostreid herpesvirus 1 in the Pacific oyster Crassostrea gigas.
Authors: Martenot, C and Oden, E and Travaillé, E and Malas, J P and Houssin, M
Journal: Journal of virological methods (2010): 86-9
Effect of perinatal short-course zidovudine on the clinical and virological manifestations of HIV-1 subtype E infection in infants.
Authors: Sutthent, Ruengpung and Chokephaibulkit, Kulkanya and Piyasujabul, Daorung and Vanprapa, Nirun and Roogpisuthipong, Anuwat and Chaisilwatana, Pongsakdi
Journal: Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology (2002): 47-56