All-in-One qPCR mix试剂与基因特异引物在通用的qPCR反应条件下配套使用,即可获得可靠的实验数据。两种产品经共同开发并进行一体化优化,表现出了如您所期望的、无需任何高成本的全范围优势。
- 均一化的反应条件缩短实验设计时间,更早提交论文
- 高扩增效率和灵敏度(甚至是低丰度基因)意味着每次实验中的可靠定量
- 无非特异性扩增和二聚体,确保可重复性和数据可靠性
All-in-One qPCR mix使用高保真热启动酶、经过优化的反应缓冲液和高质量dNTPs,能够使低丰度RNA(cDNA)或DNA进行特异的、灵敏的扩增。
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扩增曲线 | 熔解曲线 | 标准曲线 |
图1.用梯度稀释的质粒DNA(5×106至5个分子)做标准曲线来评估All-in-One™ qPCR Mix的扩增效率和检测灵敏度。如扩增曲线和熔解曲线所示,All-in-One™ qPCR Mix可扩增低至5个分子的模板(质粒DNA)说明了扩增和检测的高灵敏度。
All-in-One 编码基因验证引物 
使用GeneCopoeia精确的qPCR试剂产品可消除没玩没了的调整和优化过程。只需选择All-in-One验证引物即可开始。
All-in-One人、小鼠或大鼠的qPCR反应特异引物由专利算法设计,并为准确定量而进行了验证。验证结果包括熔解曲线,以保证扩增正确靶DNA序列(见图2.)。
与基于SYBR® Green I的All-in-One qPCR Mix试剂配套使用,All-in-One验证引物在qPCR定量实验中表现出可靠、可重复的高效性。
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扩增曲线 | |
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熔解曲线 | 验证结果的琼脂糖电泳 |
图2. 45对基因特异引物经过实验验证均产生单一扩增产物,融解曲线均呈单一锐锋,电泳条带均呈单一条带,且与目标基因相符。由10种人组织(肺、肝、睾丸、卵巢、脾、脑、胎盘、胰、心和乳腺)混合样本的总RNA经逆转录形成的cDNA作为引物验证的模板。qPCR反应由终浓度0.2 uM的引物和All-in-One qPCR Mix配合完成。使用Bio-Rad iQ5 qPCR仪器,在95°C预变性10分钟后,紧接着40个PCR循环(95°C变性10秒,60°C退火20秒钟,72°C延伸15秒),最后一个循环后设置72至95°C的熔解曲线检测程序。扩增曲线、熔解曲线和琼脂糖凝胶电泳图如图所示(验证的阳性结果见奇数泳道,阴性对照结果(NTC)见偶数泳道)。
注释:1.以上无非特异性扩增和二聚体的验证结果均建立于本公司配套试剂产品和反应体系之上。2.以上验证引物包括人、小鼠和大鼠,如需其他物种请咨询。
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- Wang, Y. et al (2013) Mechanical compaction directly modulates the dynamics of bile canaliculi formation. Integrative Biology, 2013,5, 390-40. [All-in-One™ First-strand cDNA synthesis kit].
- Xue, P. et al (2013) IGF1 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells Derived from Rat Bone Marrow by Increasing TAZ Expression. Biochemical and Biophysical Research Communications, online 5 March 2013. [All-in-One™ qPCR Mix].
- Zhou, H. et al (2013) TGF-β1 alters microRNA profile in human gastric cancer cells. Chinese Journal of Cancer, 2013 February; 25(1): 102–111. [SYBR Green Mix].
- Villanueva, J.A. et al (2013) Resveratrol Potentiates Effect of Simvastatin on Inhibition of Mevalonate Pathway in Human Endometrial Stromal Cells. Journal of Clinical Endocrinology & Metabolism, March 1, 2013 vol. 98 no. 3 E455-E462. [Primer for FPPT].
- Wang, K. et al (2013) Molecular Mechanisms Underlying the In Vitro Anti-Inflammatory Effects of a Flavonoid-Rich Ethanol Extract from Chinese Propolis (Poplar Type). Evidence-Based Complementary and Alternative Medicine, Volume 2013 (2013), Article ID 127672, 11 pages. [All-in-One™ First-strand cDNA synthesis kit].
- Li, C. et al (2013) Molecular cloning and characterization of SoNCED, a novel gene encoding 9-cis-epoxycarotenoid dioxygenase from sugarcane (Saccharum officinarum L.). Genes & Genomics, Volume 35, Issue 1, pp 101-109. [SYBR All-in-One™ qPCR Mix].
- Zhao, X. et al (2013) Identification and characterization of microRNAs from wheat (Triticum aestivum L.) under phosphorus deprivation. Journal of Plant Biochemistry and Biotechnology, January 2013, Volume 22, Issue 1, pp 113-123. [All-in-One™ First-strand cDNA synthesis kit].
- Jin, F. et al (2013) A Correlation between Altered O-GlcNAcylation, Migration and with Changes in E-cadherin Levels in Ovarian Cancer Cells. Experimental Cell Research, online 22 March 2013. [All-in-One™ First-strand cDNA synthesis kit].
- Han, X. et al. (2012) GHGKHKNK Octapeptide (P-5m) Inhibits Metastasis of HCCLM3 Cell Lines via Regulation of MMP-2 Expression in in Vitro and in Vivo Studies. Molecules, 17(2), 1357-1372; doi:10.3390/molecules17021357. [SYBR Green I reaction mix].
- Du, J. et al. (2012) Biodegradable nanoparticles of mPEG-PLGA-PLL triblock copolymers as novel non-viral vectors for improving siRNA delivery and gene silencing. International Journal of Molecular Sciences, 13.1 (2012): 516-533. [All-in-One™ First-strand cDNA synthesis kit].