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.
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