Figure 1. Forget-Me-Not? allows you to see at a glance whether you’ve added reaction components to your tubes or wells. Left: 1X Forget-Me-Not? Master Mix; Right: Forget-Me-Not? Master Mix after addition of Forget-Me-Not? Template Buffer.
EvaGreen?染料是Biotium公司研發的新一代DNA結合染料,它同時適合于實時定量PCR以及高分辨率熔解曲線分析(HRM)。這種染料通過一種被稱為“按要求釋放”的全新機制,選擇性結合雙鏈DNA,從而大大降低了染料對PCR反應的抑制作用。EvaGreen?染料的另一個重要的特性是安全、無毒。目前商業化的核酸染料大都會誘導基因的突變,而且很少做過完整的毒性研究,這對使用者的健康存在著巨大的潛在危害。比如在PCR實驗中常用的SYBR? Green I染料,有文獻報道其比EB具有更大的環境毒性(Ohta, et al. Mutation Research, 492,91-97(2001)).,雖然SYBR? Green I 本身誘變性很弱,但由于它可能在活細胞中抑制正常DNA的修復機制,使其有誘變增強的作用??紤]到PCR的廣泛使用,核酸染料的安全性應該被足夠重視,基于以上的理念,Biotium的科學家研發出了新一代的核酸染料EvaGreen?。EvaGreen?由于降低了細胞膜穿透性,因而比SYBR? Green I更加安全(圖-2)。獨立實驗室的測試結果顯示,EvaGreen?既沒有誘變性也沒有細胞毒性,廢棄物可直接倒入下水 道。完整的安全性報告請下載EvaGreen? Dye Safety Report。
Figure 2. Comparison of GAPDH fast-cycling real time qPCR assay using Forget-Me-Not? qPCR Master Mix, Biotium’s Fast EvaGreen? qPCR Master Mix, or Qiagen’s QuantiNova? SYBR? Green qPCR Master Mix. A. Raw fluorescence data. EvaGreen?-based master mixes yield higher fluorescence signal compared to the SYBR? Green-based master mix. B. Data plotted on logarithmic scale. Forget-Me-Not? performs as well or better than other PCR master mixes.
EvaGreen? Dye and Cheetah? HotStart Taq DNA Polymerase are covered under US and international patents. QuantiNova? is a registered trademark of Qiagen Group. SYBR? Green is a registered trademark of Molecular Probes, Inc./Life Technologies. *HRM? is a registered trademark of Idaho Technologies, Inc./BioFire Defense, LLC and its use may require a license.