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Q4ever Green核酸染料 *2000X DMSO 溶液*

貨號17608存儲條件 在零下15度以下保存, 避免光照
規格100 ul價格1740
Ex (nm)503Em (nm)527
分子量溶劑Water
產品詳細介紹


簡要概述

實時聚合酶鏈反應(real-time PCR),也稱為定量聚合酶鏈反應(qPCR),是一種基于聚合酶鏈反應(PCR)的流行的分子生物學實驗室技術。它可以在PCR期間(即實時)檢測目標DNA分子的擴增情況,而不是像常規PCR那樣在結束時進行檢測。實時PCR可以定量(定量實時PCR)和半定量(即高于或低于一定數量的DNA分子)(半定量實時PCR)使用。實時PCR中有兩種檢測PCR產物的常用方法,包括(1)結合任何雙鏈DNA的非特異性熒光染料; (2)由寡核苷酸組成的序列特異性DNA探針,所述寡核苷酸用熒光報告物標記,僅在探針與其互補序列雜交后才允許檢測。對于第一種方法,對DNA結合染料進行PCR的實時檢測有兩個要求,即(a)與雙鏈DNA結合時增強熒光;(b)對PCR的抑制作用最小。 SYBR Green主要用于各種qPCR應用中。我們最近開發了新一代的SYBR Green Q4ever Green,用于解決SYBR Green的一些局限性,例如酶抑制。 Q4ever Green可以在PCR中使用,幾乎沒有PCR抑制作用,并提高了靈敏度。 Q4ever Green可用于檢測任何雙鏈DNA序列的擴增。假設您的PCR引物設計合理且反應特性良好,則無需探針,可以減少測定設置和運行成本。作為SYBR Green,主要缺點是它可能產生假陽性信號。即因為Q4ever Green染料與任何雙鏈DNA結合。它也可以與非特異性雙鏈DNA序列結合。設計良好的引物不擴增非靶序列,并進行熔解曲線分析,這一點極為重要。



產品說明書

實驗方案

儲存在-20°C,避光。 按建議存放時,產品自收到之日起至少穩定12個月。

 

工作溶液配制

Q4ever Green工作溶液(50X)
使用水或TE緩沖液稀釋2000X Q4ever Green儲備溶液以制成50X Q4ever Green儲備溶液。

 

操作步驟

建議使用以下協議。 如果需要,可以調整方案以達到最佳效果。

設置PCR反應如下:
5 µL 10X聚合酶緩沖液(不含鎂)
2.5 µL的50 mM MgCl2
2 µL 50X Q4ever Green工作溶液
2 µL的5 mM dUTP
1-5個單位的DNA聚合酶
加入所需的cDNA
每個引物100-1000 nM(正向和反向引物的終濃度)
用dH2O將最終體積調整為50 µL。
在熱循環熒光計上執行實時PCR并記錄熒光信號。

 

參考文獻

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

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

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

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

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

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

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

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

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

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



說明書
Q4ever Green核酸染料 *2000X DMSO 溶液*.pdf


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