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AAT Bioquest

NED Dye qPCR Calibration Solution *10,000X*

ABI 7500 FAST system NED dye spectrum.
ABI 7500 FAST system NED dye spectrum.
ABI 7500 FAST system NED dye spectrum.
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Physical properties
Molecular weightN/A
SolventWater
Spectral properties
Absorbance (nm)544
Extinction coefficient (cm -1 M -1)740001
Excitation (nm)545
Emission (nm)567
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure

OverviewpdfSDSpdfProtocol


Molecular weight
N/A
Absorbance (nm)
544
Extinction coefficient (cm -1 M -1)
740001
Excitation (nm)
545
Emission (nm)
567
Real-time thermal cyclers require calibration to generate a spectral profile for the fluorescence signal. Since instruments can differ in fluorescence detection sensitivity, the concentrations needed to calibrate any dye, such as NED, will depend on the device used. The AAT Bioquest NED dye qPCR Calibration Solution supplied as a 10,000X concentrate is compatible with a wide variety of qPCR instruments, including the ABI7500 Fast, QuantStudio™, and ViiA™ 7 systems. Calibration and verification should be run every six months or as needed. Please refer to the instrument maintenance guide for complete instructions on use.

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

This generic protocol is suitable for the ABI7500 FAST qPCR instrument. For complete instructions on use, refer to the maintenance guide for your instrument.

Prepare a NED calibration plate:
  1. Prepare a NED Dye qPCR Calibration working solution by mixing 1 µL of the 10,000X NED Dye qPCR Calibration Solution with 10 mL of nuclease-free water or qPCR buffer. Note the final dye concentration must be optimized based on the calibration requirements for your instrument. Refer to the manufacturer’s maintenance guide for detailed instructions.
  2. Add 20 µL of the NED Dye qPCR Calibration working solution to all wells of a reaction plate. Note the volume may vary depending on your instrument. Refer to the manufacturer’s maintenance guide for detailed instructions.
  3. Seal the plate with an optical adhesive cover.
  4. Centrifuge the plate for 2 minutes at < 1500 rpm.
  5. Check that the liquid in each well of the plate is at the bottom of the well. If not, centrifuge the plate again at a slightly higher rpm and for a longer period of time.
  6. For detailed calibration instructions, refer to the maintenance guide for your instrument. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Absorbance (nm)544
Extinction coefficient (cm -1 M -1)740001
Excitation (nm)545
Emission (nm)567

Images


References


View all 36 references: Citation Explorer
Detection of nitric oxide radical and determination of its scavenging activity by antioxidants using spectrophotometric and spectrofluorometric methods.
Authors: Can, Ziya and Keskin, Büşra and Üzer, Ayşem and Apak, Reşat
Journal: Talanta (2022): 122993
An In-Vitro Evaluation of Microleakage in Resin-Based Restorative Materials at Different Time Intervals.
Authors: Bilgrami, Afreen and Alam, Mohammad Khursheed and Qazi, Fazal Ur Rehman and Maqsood, Afsheen and Basha, Sakeenabi and Ahmed, Naseer and Syed, Kausar Ali and Mustafa, Mohammed and Shrivastava, Deepti and Nagarajappa, Anil Kumar and Srivastava, Kumar Chandan
Journal: Polymers (2022)
DNA degradation in human teeth exposed to thermal stress.
Authors: Lozano-Peral, Diego and Rubio, Leticia and Santos, Ignacio and Gaitán, María Jesús and Viguera, Enrique and Martín-de-Las-Heras, Stella
Journal: Scientific reports (2021): 12118
Development of a gold nanoparticles-based colorimetric sensor for the indirect determination of ammonium dinitramide and tetryl.
Authors: Mamatioğlu, Fatma and Üzer, Ayşem and Erçağ, Erol and Apak, Reşat
Journal: Talanta (2021): 122187
Fluorescent Probe for Ag+ Detection Using SYBR GREEN I and C-C Mismatch.
Authors: Zhou, Xiaohong and Memon, Abdul Ghaffar and Sun, Weiming and Fang, Fang and Guo, Jinsong
Journal: Biosensors (2020)
Comparison of Retrograde, Primary and Secondary Bonding Materials with Tooth Substance.
Authors: Khan, Tabinda Nawaz and Abidi, Syed Yawar Ali
Journal: Journal of the College of Physicians and Surgeons--Pakistan : JCPSP (2018): 9-12
Simultaneous detection and quantification of 19 diarrhea-related pathogens with a quantitative real-time PCR panel assay.
Authors: Wongboot, Warawan and Okada, Kazuhisa and Chantaroj, Siriporn and Kamjumphol, Watcharaporn and Hamada, Shigeyuki
Journal: Journal of microbiological methods (2018): 76-82
Micromechanical Intervention in Sandwich Restoration.
Authors: Khan, Tabinda Nawaz and Ali Abidi, Syed Yawar and Nawaz Khan, Khush Bakht and Ahmed, Shahbaz and Rehman Qazi, Fazal Ur and Saeed, Noureen
Journal: Journal of the College of Physicians and Surgeons--Pakistan : JCPSP (2015): 781-4
4-Aminothiophenol functionalized gold nanoparticle-based colorimetric sensor for the determination of nitramine energetic materials.
Authors: Üzer, Ayşem and Can, Ziya and Akın, Ilknur and Erçağ, Erol and Apak, Reşat
Journal: Analytical chemistry (2014): 351-6
Role of caffeic Acid on collagen production in nasal polyp-derived fibroblasts.
Authors: Chung, Seung Won and Park, Il-Ho and Hong, Sung-Moon and Cho, Jung-Sun and Moon, Jun-Hyeok and Kim, Tae Hoon and Lee, Heung-Man
Journal: Clinical and experimental otorhinolaryngology (2014): 295-301