Q4ever™ Green *1250X DMSO Solution*
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
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International | See distributors |
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Shipping | Standard overnight for United States, inquire for international |
Physical properties
Solvent | Water |
Spectral properties
Excitation (nm) | 503 |
Emission (nm) | 527 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12171501 |
Overview | ![]() ![]() |
See also: Kinases, Genome Editing & CRISPR, Polymerase Chain Reaction (PCR), Real-Time PCR (qPCR), Reverse Transcription PCR (RT-PCR)
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.
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
Set up the PCR reaction as follows:
5 µL of 10X polymerase buffer without magnesium2.5 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.Product Family
Name | Excitation (nm) | Emission (nm) |
Indocyanine Green *CAS 3599-32-4* | 789 | 813 |
Helixyte™ Green *20X Aqueous PCR Solution* | 498 | 522 |
Helixyte™ Green *10,000X Aqueous PCR Solution* | 498 | 522 |
Thiolite™ Green | 505 | 524 |
CytoTell™ Green | 510 | 525 |
LysoBrite™ Green | 501 | 510 |
Droplite™ Green | 421 | 521 |
Images
References
View all 50 references: Citation Explorer
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
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
Authors: Ajayi, A and Jolaiya, T and Smith, S I
Journal: BMC research notes (2021): 90
Quantitative Assessment of Parental Somatic Mosaicism for Copy-Number Variant (CNV) Deletions.
Authors: Liu, Qian and Grochowski, Christopher M and Bi, Weimin and Lupski, James R and Stankiewicz, Paweł
Journal: Current protocols in human genetics (2020): e99
Authors: Liu, Qian and Grochowski, Christopher M and Bi, Weimin and Lupski, James R and Stankiewicz, Paweł
Journal: Current protocols in human genetics (2020): e99
Detection of Phytophthora infestans by Loop-Mediated Isothermal Amplification, Real-Time LAMP, and Droplet Digital PCR.
Authors: Ristaino, Jean B and Saville, Amanda C and Paul, Rajesh and Cooper, Donald C and Wei, Qingshan
Journal: Plant disease (2020): 708-716
Authors: Ristaino, Jean B and Saville, Amanda C and Paul, Rajesh and Cooper, Donald C and Wei, Qingshan
Journal: Plant disease (2020): 708-716
Portable and accurate diagnostics for COVID-19: Combined use of the miniPCR thermocycler and a well-plate reader for SARS-CoV-2 virus detection.
Authors: González-González, Everardo and Trujillo-de Santiago, Grissel and Lara-Mayorga, Itzel Montserrat and Martínez-Chapa, Sergio Omar and Alvarez, Mario Moisés
Journal: PloS one (2020): e0237418
Authors: González-González, Everardo and Trujillo-de Santiago, Grissel and Lara-Mayorga, Itzel Montserrat and Martínez-Chapa, Sergio Omar and Alvarez, Mario Moisés
Journal: PloS one (2020): e0237418
Development of an EvaGreen-based real-time PCR assay for detection of Aleutian mink disease virus.
Authors: Li, Li and Hu, Zhe and Sun, Jinhui and Guo, Kui and Chu, Xiaoyu and Wang, Xiaojun and Lu, Yixin
Journal: Journal of virological methods (2020): 113751
Authors: Li, Li and Hu, Zhe and Sun, Jinhui and Guo, Kui and Chu, Xiaoyu and Wang, Xiaojun and Lu, Yixin
Journal: Journal of virological methods (2020): 113751
A handheld continuous-flow real-time fluorescence qPCR system with a PVC microreactor.
Authors: Shi, Bing and Li, Yuanming and Wu, Di and Wu, Wenming
Journal: The Analyst (2020): 2767-2773
Authors: Shi, Bing and Li, Yuanming and Wu, Di and Wu, Wenming
Journal: The Analyst (2020): 2767-2773
Epidemiological investigation of feline infectious peritonitis in cats living in Harbin, Northeast China from 2017 to 2019 using a combination of an EvaGreen-based real-time RT-PCR and serum chemistry assays.
Authors: Guan, Xueting and Li, Hua and Han, Meijing and Jia, Shuo and Feng, Baohua and Gao, Xuwen and Wang, Zhuo and Jiang, Yanping and Cui, Wen and Wang, Li and Xu, Yigang
Journal: Molecular and cellular probes (2020): 101495
Authors: Guan, Xueting and Li, Hua and Han, Meijing and Jia, Shuo and Feng, Baohua and Gao, Xuwen and Wang, Zhuo and Jiang, Yanping and Cui, Wen and Wang, Li and Xu, Yigang
Journal: Molecular and cellular probes (2020): 101495
Development of an EvaGreen based real-time RT-PCR assay for rapid detection, quantitation and diagnosis of goose calicivirus.
Authors: Lin, Su and Zhang, Shizhong and Wang, Shao and Xie, Kaichun and Jiang, Dandan and Xiao, Shifeng and Chen, Xiuqin and Chen, Shaoying
Journal: Molecular and cellular probes (2020): 101489
Authors: Lin, Su and Zhang, Shizhong and Wang, Shao and Xie, Kaichun and Jiang, Dandan and Xiao, Shifeng and Chen, Xiuqin and Chen, Shaoying
Journal: Molecular and cellular probes (2020): 101489
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
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
Application notes
FAQ
Does ethidium bromide stain RNA?
Are Cell Navigator® Cell Plasma Membrane Staining Kits suitable for cell culture medium samples?
Are mRNAs found in prokaryotes differ from those of eukaryotes?
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Are Cell Navigator® Cell Plasma Membrane Staining Kits suitable for cell culture medium samples?
Are mRNAs found in prokaryotes differ from those of eukaryotes?
Are spliceosomes associated with any diseases?
Are there any alternatives for ethidium bromide in agarose gels?