EvoGreen Dye (20× in water) 是一种饱和双链DNA结合荧光染料,与SYBR Green I 有相似的光谱特性 (Ex 495 nm,Em 520 nm),但具有以下三个优势:
1. EvoGreen Dye通过“按需求释放”机制结合DNA,灵敏度高,对Bst DNA聚合酶的抑制远小于SYBR Green I,更适合环介导等温扩增 (LAMP)反应。使用EvoGreen荧光染料可以不开盖检测LAMP扩增产物, 避免了开盖电泳检测带来的扩增子气溶胶污染,结果更为可靠。
2. EvoGreen Dye稳定性极强,在正常的储存、稀释加样和DNA扩增过程中不会被破坏。缓冲溶液中的染料可以安全地储存在室温或冰箱里,也可以反复冻融。
3. EvoGreen Dye既没有诱变性也几乎不能穿透细胞膜,安全性高。
使用方法
按照正常操作流程配制扩增反应体系,加入1/20体积的EvoGreen Dye,使最终反应体系中的EvoGreen Dye浓度为1×,其他操作步骤和仪器设备均无需改变。
EvoGreen Dye (20× in water) is a saturating fluorescent dye for double-stranded DNA binding, with spectral properties similar to SYBR Green I (Ex 495 nm, Em 520 nm), but offers the following three advantages:
1. EvoGreen Dye binds to DNA through a "release-on-demand" mechanism with high sensitivity. Its inhibition of Bst DNA polymerase is much weaker than that of SYBR Green I, making it more suitable for loop-mediated isothermal amplification (LAMP) reactions. Using EvoGreen fluorescent dye allows LAMP amplification products to be detected without opening the reaction tube, avoiding amplicon aerosol contamination caused by open-lid electrophoresis detection and yielding more reliable results.
2. EvoGreen Dye exhibits excellent stability and is not degraded during routine storage, dilution, pipetting and DNA amplification. The dye in buffer solution can be safely stored at room temperature or in a refrigerator, and can also withstand repeated freeze-thaw cycles.
3. EvoGreen Dye is non-mutagenic and barely penetrates cell membranes, ensuring high biosafety.
Protocol
Prepare the amplification reaction mixture following standard procedures. Add EvoGreen Dye at a volume ratio of 1/20 to achieve a final concentration of 1× in the reaction system. No changes to other operating steps or instruments are required.