EveGreen (20×in water)

Cat.No. LM603001
Size500 µL/1 mL
Storage and shippingTransport on ice packs. Store away from light at 4°C. Shelf life is 24 months.

PDF Download: LM603001

 

Product description

EveGreen is a novel DNA-binding dye used in real-time quantitative PCR (qPCR). This dye is designed based on “release-on-demand” DNA binding technology. Structurally, it consists of two monomeric dye molecules linked by a flexible arm to form a dimer. In the absence of DNA, this structure adopts a non-fluorescent ring-shaped conformation. However, in the presence of DNA, it undergoes a conformational change and binds to DNA, emitting strong green fluorescence.

Compared to other fluorescent dyes on the market such as FAM and SYBR Green I, EveGreen possesses several superior characteristics:

  • Extremely high sensitivity: When used at the recommended concentration, it provides a stronger PCR amplification signal compared to SYBR Green I.
  • Minimal PCR inhibition, making it highly suitable for multiplex PCR and HRM analysis: The “release-on-demand” DNA binding technology significantly reduces PCR inhibition compared to SYBR Green I. Therefore, qPCR experiments using EveGreen can utilize rapid PCR cycling conditions. Additionally, EveGreen can be used at higher concentrations, resulting in significantly stronger amplification signals compared to SYBR Green I. The higher concentration of EveGreen also eliminates dye redistribution defects, allowing it to be used for both multiplex PCR and high-resolution melt curve analysis.
  • Exceptional stability: It remains stable throughout normal storage, handling, and PCR processes. The dye in buffer solution can be safely stored at room temperature or in a refrigerator, and can undergo repeated freeze-thaw cycles.
  • Excellent safety profile: Cell membrane permeability tests have shown that EveGreen has minimal cell membrane penetration and is highly safe. It exhibits no mutagenicity or cytotoxicity.
  • Superior compatibility: It has a similar spectrum to SYBR Green I and is compatible with various leading brands of qPCR instruments. Substituting EveGreen for SYBR Green I does not require any changes to current operating procedures or instrument settings.

Moreover, EveGreen is currently the only dye available for use in droplet digital PCR systems, and it demonstrates outstanding performance in microfluidic PCR, isothermal amplification, capillary electrophoresis DNA quantification, and cell viability assays. This product is provided in the form of a 20× EveGreen aqueous solution.

 

qPCR reaction System

Table 1: According to the table below, calculate the amounts of each component based on the prepared system.

ReagentFinal concentration in the mixture
ddH2OAdjust to final volume
Taq polymerase buffer without magnesium
dNTP0.2 mM each
MgCl22.5 mM
EveGreen
Taq DNA polymerase1-5 units per reaction
Each of primers0.1-1 μM

[Note]:

  1. Please add or omit the appropriate concentration of the reference dye ROX according to the type of instrument used.
  2. This system needs to be appropriately optimized based on the type of Taq enzyme used.
  3. According to the experimental requirements, sequentially add water, Taq polymerase buffer, dNTPs, MgCl2, EveGreen, Taq polymerase, and primers into sterile centrifuge tubes placed on ice to prepare the qPCR reaction system (1×Master mix).
  4. Dispense an appropriate volume of 1×Master mix into sterile PCR tubes, and add DNA (50 ng/reaction).
  5. Select the appropriate amplification program according to the type of instrument.

 

Notes

  1. When performing qPCR, ensure to set positive and negative controls.
  2. For your safety and health, wear a lab coat and disposable gloves during operation.
  3. This product is for research use only!

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