Applied BioProbes公司ZX推出了一系列核酸荧光染料产品作为EB替代品,具有安全、灵敏度高以及产品性能稳定等优点。 GreenView™, GreenView™ Plus, GreenView™ Ultra, and RedView™是Applied BioProbes公司ZX研发具有安全无毒,灵敏度高,稳定性好的EB替代品, 使用方法同EB,可用于前染法和后染法,条带整齐不弯曲。GreenView™, GreenView™ Plus, and GreenView™ Ultra可在紫外灯或蓝光仪下观察发绿色荧光;RedView™可在紫外灯下观察发红色荧光。 核酸电泳是核酸研究的重要手段之一,其中溴化乙啶(EB)因其低廉的价格是分子生物学中使用和最为广泛的核酸电泳染色试剂。但是EB是一种强诱变剂,具有高致癌性,其废弃液必须进行净化处理,而且EB的检测灵敏度有限以及背景信号较高,目前正逐渐被灵敏度高以及安全可靠的新型核酸荧光染料所取代。
产品特点:
安全无毒,无致突变
稳定性好
灵敏度高,条带整齐
Figure 1. Gel stain results with GreenView™, GreenView™ Plus, and GreenView™ Ultra (Left), and comparison results with SYBR Safe™ and GelGreen™ (Right). Two-fold serial dilutions of a 1 kb DNA ladder (NEB, Cat # N0468S) were loaded onto each gel in 3 lanes in the amounts of 25 ng, 50 ng and 100 ng (left to right), then post-stained with GreenView™, GreenView™ Plus, GreenView™ Ultra, SYBR Safe™ and GelGreen™, respectively. Results show GreenView™ has similar sensitivity with SYBR Safe™ and GelGreen™, GreenView™ Plus and GreenView™ Ultra have much better sensitivity.
Figure 2. Gel stain results with RedView™, and GelRed™. Two-fold serial dilutions of a 1 kb DNA ladder (NEB, Cat # N0468S) were loaded onto each gel in 3 lanes in the amounts of 25 ng, 50 ng and 100 ng (left to right), then post-stained with RedView™, and GelRed™, respectively. Results show RedView™ has similar sensitivity with GelRed™. 此外,GreenView™, GreenView™ Plus, and RedView™ 具有较低的细胞膜通透性,不易与细胞内DNA结合,安全性优于SYBR Safe。
Figure 3. Cell permeability tests with SYBR Safe, GreenView, GreenView Plus, and RedView. Hela cells were incubated at 37℃ with 1X SYBR Safe, GreenView, GreenView Plus, and RedView for 30 min. GreenView Plus and RedView could not penetrate cell membranes to bind DNA in living cells, and were much safer than SYBR Safe, which rapidly entered cells and stained nuclei.
SafeGreen™ 和 SafeRed™ Loading Dye 含 6 × DNA 上样缓冲液中及两类常用的示踪染料 (Xylene Cyanol FF 和 Bromophenol Blue),用以监测电泳期间琼脂糖凝胶条带(呈现紫色和蓝色两个条带)迁移情况。DNA条带可通过 LED 或紫外光直接观察,无需前染和后染步骤。
Figure 4. Gel staining results with SafeGreen™ and SafeRed™ premixed with DNA ladder. 1 kb DNA Ladder (Cat# D001) was mixed with SafeGreen™ or SafeRed™ at a 5:1 ratio, then loaded onto each well. Gels were directly imaged on a UV transilluminator. SafeGreen™ 和 SafeRed™ 无法穿透细胞膜进入活细胞,从而降低了细胞毒性(Figure 5)。
Figure 5. Cell permeability tests with SYBR Safe™, SafeGreen™, and SafeRed™. Hela cells were incubated at 37℃ with 1X SYBR Safe™, SafeGreen™, and SafeRed™ for 30 min. SafeGreen™ and SafeRed™ could not penetrate cell membranes to bind DNA in living cells, and were much safer than SYBR Safe™, which rapidly entered cells and stained nuclei.