All-in-One qPCR mix試劑與基因特異引物在通用的qPCR反應條件下配套使用,即可獲得可靠的實驗數據。兩種產品經共同開發并進行一體化優化,表現出了如您所期望的、無需任何高成本的全范圍優勢。
- 均一化的反應條件縮短實驗設計時間,更早提交論文
- 高擴增效率和靈敏度(甚至是低豐度基因)意味著每次實驗中的可靠定量
- 無非特異性擴增和二聚體,確保可重復性和數據可靠性
All-in-One qPCR mix使用高保真熱啟動酶、經過優化的反應緩沖液和高質量dNTPs,能夠使低豐度RNA(cDNA)或DNA進行特異的、靈敏的擴增。
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擴增曲線 | 熔解曲線 | 標準曲線 |
圖1.用梯度稀釋的質粒DNA(5×106至5個分子)做標準曲線來評估All-in-One™ qPCR Mix的擴增效率和檢測靈敏度。如擴增曲線和熔解曲線所示,All-in-One™ qPCR Mix可擴增低至5個分子的模板(質粒DNA)說明了擴增和檢測的高靈敏度。
All-in-One 編碼基因驗證引物 
使用GeneCopoeia精確的qPCR試劑產品可消除沒玩沒了的調整和優化過程。只需選擇All-in-One驗證引物即可開始。
All-in-One人、小鼠或大鼠的qPCR反應特異引物由專利算法設計,并為準確定量而進行了驗證。驗證結果包括熔解曲線,以保證擴增正確靶DNA序列(見圖2.)。
與基于SYBR® Green I的All-in-One qPCR Mix試劑配套使用,All-in-One驗證引物在qPCR定量實驗中表現出可靠、可重復的高效性。
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擴增曲線 | |
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熔解曲線 | 驗證結果的瓊脂糖電泳 |
圖2. 45對基因特異引物經過實驗驗證均產生單一擴增產物,融解曲線均呈單一銳鋒,電泳條帶均呈單一條帶,且與目標基因相符。由10種人組織(肺、肝、睪丸、卵巢、脾、腦、胎盤、胰、心和乳腺)混合樣本的總RNA經逆轉錄形成的cDNA作為引物驗證的模板。qPCR反應由終濃度0.2 uM的引物和All-in-One qPCR Mix配合完成。使用Bio-Rad iQ5 qPCR儀器,在95°C預變性10分鐘后,緊接著40個PCR循環(95°C變性10秒,60°C退火20秒鐘,72°C延伸15秒),最后一個循環后設置72至95°C的熔解曲線檢測程序。擴增曲線、熔解曲線和瓊脂糖凝膠電泳圖如圖所示(驗證的陽性結果見奇數泳道,陰性對照結果(NTC)見偶數泳道)。
注釋:1.以上無非特異性擴增和二聚體的驗證結果均建立于本公司配套試劑產品和反應體系之上。2.以上驗證引物包括人、小鼠和大鼠,如需其他物種請咨詢。
- Wang, Y.Z. et al (2013) Macrophage migration inhibitory factor is necessary for the Lipo-oligosaccharide-induced response by modulation of Toll-like receptor 4 in monocytes from GBS… Journal of Neuroimmunology, online 12 February 2013. [All-in-One™ First-strand cDNA synthesis kit].
- 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].