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Q4ever™ Green *1250X DMSO Solution*

A comparison of the relative fluorescence signal from qPCR reactions performed with Q4ever<sup>TM</sup> Green and SYBR<sup>TM</sup> Green. Q4ever<sup>TM</sup> Green has much brighter signal than SYBR<sup>TM</sup> Green.
A comparison of the relative fluorescence signal from qPCR reactions performed with Q4ever<sup>TM</sup> Green and SYBR<sup>TM</sup> Green. Q4ever<sup>TM</sup> Green has much brighter signal than SYBR<sup>TM</sup> Green.
Quantitative PCR results targeting GAPDH with an input of 100 ng–0.00001 ng cDNA was performed using Q4ever™ Green *1250X DMSO Solution* (Cat No. 17608) and a Fast Advanced Master Mix on an Applied Biosystems® 7500 FAST Real-Time PCR System.
Ordering information
Price ()
Catalog Number17608
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
Physical properties
SolventWater
Spectral properties
Excitation (nm)503
Emission (nm)527
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
UNSPSC12171501

OverviewpdfSDSpdfProtocol


Excitation (nm)
503
Emission (nm)
527
A real-time polymerase chain reaction (real-time PCR), also known as quantitative polymerase chain reaction (qPCR), is a popular laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR (i.e., in real time), not at its end, as in conventional PCR. Real-time PCR can be used quantitatively (quantitative real-time PCR) and semi-quantitatively (i.e., above/below a certain amount of DNA molecules) (semi-quantitative real-time PCR). There are two common methods for the detection of PCR products in real-time PCR, including (1) non-specific fluorescent dyes that bind any double-stranded DNA; and (2) sequence-specific DNA probes consisting of oligonucleotides that are labelled with a fluorescent reporter, which permits detection only after hybridization of the probe with its complementary sequence. For the first method, there are two requirements for a DNA binding dye for real-time detection of PCR, i.e., (a). enhanced fluorescence when bound to double-stranded DNA; and (b). minimal inhibition of PCR. SYBR Green is predominantly used in a variety of qPCR applications. We have recently developed Q4ever™Green, a newer generation of SYBR Green, to address some limitations with SYBR Green, e.g., enzyme inhibition. Q4ever™Green permits the use of the Q4ever™Green in PCR with little PCR inhibition and enhanced sensitivity. Q4ever™Green can be used to monitor the amplification of any double-stranded DNA sequence. No probe is required, which can reduce assay setup and running costs, assuming that your PCR primers are well designed and your reaction is well characterized. As SYBR Green, the primary disadvantage is that it may generate false positive signals; i.e., because the Q4ever™Green dye binds to any double-stranded DNA. It can also bind to nonspecific double-stranded DNA sequences. It is extremely important to have well-designed primers that do not amplify non-target sequences, and that melt curve analysis be performed.

Example protocol


AT A GLANCE

Storage and Handling
Store at -20 °C, protected from light. Product is stable for at least 12 months from the date of receipt when stored as recommended.

Safety
We advise researchers to follow universal laboratory safety precautions when handling Q4ever Green dye.

PREPARATION OF WORKING SOLUTION

Q4ever Green working solution (50X)
Dilute the 1250X Q4ever Green stock solution to make a 50X Q4ever Green stock solution using water or TE buffer.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol is recommended. Adjust the protocol if needed to achieve optimal results.

Set up the PCR reaction as follows:
5 µL of 10X polymerase buffer without magnesium
2.5 µL of 50 mM MgCl2
2 µL of 50X Q4ever Green working solution
2 µL of 5 mM dUTP
1-5 units of DNA polymerase
Desired amount of cDNA
100-1000 nM each of primers (Final concentrations of Forward and Reverse primers)
Adjust the final volume to 50 µL with dH2O

Perform real-time PCR
Perform real-time PCR on a thermocycling fluorometer and record the fluorescence signal.

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)503
Emission (nm)527

References


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MinION Nanopore-based detection of Clavibacter nebraskensis, the corn Goss's wilt pathogen, and bacteriomic profiling of necrotic lesions of naturally-infected leaf samples.
Authors: Xu, Renlin and Adam, Lorne and Chapados, Julie and Soliman, Atta and Daayf, Fouad and Tambong, James T
Journal: PloS one (2021): e0245333
Direct detection of Helicobacter pylori from biopsies of patients in Lagos, Nigeria using real-time PCR-a pilot study.
Authors: Ajayi, A and Jolaiya, T and Smith, S I
Journal: BMC research notes (2021): 90
High-Level Multiplexing in Digital PCR with Intercalating Dyes by Coupling Real-Time Kinetics and Melting Curve Analysis.
Authors: Moniri, Ahmad and Miglietta, Luca and Holmes, Alison and Georgiou, Pantelis and Rodriguez-Manzano, Jesus
Journal: Analytical chemistry (2020): 14181-14188
Amplification Curve Analysis: Data-Driven Multiplexing Using Real-Time Digital PCR.
Authors: Moniri, Ahmad and Miglietta, Luca and Malpartida-Cardenas, Kenny and Pennisi, Ivana and Cacho-Soblechero, Miguel and Moser, Nicolas and Holmes, Alison and Georgiou, Pantelis and Rodriguez-Manzano, Jesus
Journal: Analytical chemistry (2020): 13134-13143
A sensitive assay for dNTPs based on long synthetic oligonucleotides, EvaGreen dye and inhibitor-resistant high-fidelity DNA polymerase.
Authors: Purhonen, Janne and Banerjee, Rishi and McDonald, Allison E and Fellman, Vineta and Kallijärvi, Jukka
Journal: Nucleic acids research (2020): e87
Detection of extended-spectrum beta-lactamase cefotaxime resistance and virulence genes in Escherichia coli by duplex quantitative real-time PCR and melt curve analysis.
Authors: Aijuka, M and Buys, E M
Journal: Letters in applied microbiology (2020): 54-60
A quantitative loop-mediated isothermal amplification assay for detecting a novel goose astrovirus.
Authors: He, Dalin and Yang, Jing and Jiang, Xiaoning and Lin, Yun and Chen, Hao and Tang, Yi and Diao, Youxiang
Journal: Poultry science (2020): 6586-6592
Comprehensive Data of P53 R282 Gene Mutation with Human Papillomaviruses (HPV)-Associated Oral Squamous Cell Carcinoma (OSCC).
Authors: Ekalaksananan, Tipaya and Wongjampa, Weerayut and Phusingha, Pensiri and Chuerduangphui, Jureeporn and Vatanasapt, Patravoot and Promthet, Supannee and Patarapadungkit, Natcha and Pientong, Chamsai
Journal: Pathology oncology research : POR (2020): 1191-1199
Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection.
Authors: Falzone, Luca and Musso, Nicolò and Gattuso, Giuseppe and Bongiorno, Dafne and Palermo, Concetta Ilenia and Scalia, Guido and Libra, Massimo and Stefani, Stefania
Journal: International journal of molecular medicine (2020): 957-964
New chalcone-tethered 1,3,5-triazines potentiate the anticancer effect of cisplatin against human lung adenocarcinoma A549 cells by enhancing DNA damage and cell apoptosis.
Authors: El-Wakil, Marwa H and Khattab, Sherine N and El-Yazbi, Amira F and El-Nikhely, Nefertiti and Soffar, Ahmed and Khalil, Hosam H
Journal: Bioorganic chemistry (2020): 104393