Chai Green Dye 20X

Highlights

• A superior dye for Real-Time PCR & High Resolution Melt (HRM)
• Dramatically lower PCR inhibition compared with SYBR Green
• Open license allows free use, with no restrictions or licensing fees
• Compatible with all instruments supporting SYBR/FAM fluorophores

Description

Chai Green™ is a new DNA binding dye optimized for Real-Time PCR and High Resolution Melt (HRM) analysis. It is significantly less inhibitory than SYBR Green because it binds primarily within the minor groove of DNA. Because of its lower inhibition, it may be used in higher concentrations resulting in increased fluorescence output, making it ideal for sensitive applications like HRM. The decreased PCR inhibition also decreases detection times in isothermal amplification applications.

Chai Green is compatible with all Real-Time PCR instruments supporting SYBR or FAM fluorophores, as its fluorescence spectrum is similar to SYBR Green. Dye binding appears to be cooperative and is influenced by the dye concentration and dye-to-basepair ratio.

Chai Green Fluorescence Spectrum

The fluorescence spectrum of Chai Green™ is similar to that of SYBR Green.

Features

• Thermally stable dye designed for qPCR, melt curve, and High Resolution Melt (HRM) analysis
• Perform HRM at 1X working concentration without inhibiting PCR
• No detectable loss of fluorescence after 20 freeze/thaw cycles
• Produces lower detection times when used in isothermal amplification
• Compatible with fast PCR protocols
• No change of melt temperature when used at higher concentrations

Performance

1X Chai Green Dye: 101% Amplification Efficiency

Five ten-fold serial dilutions of a GAPDH gene fragment were used to create a PCR standard curve. A 123 bp product was amplified using 1X Chai Green concentration and Chai's 2X PCR Master Mix + Hot Start. An efficiency of 101% was obtained, with a slope of -3.328 and R₂ value of 1.000.

Chai Green Fluorescence Compared With SYBR Green I

1X Chai Green (blue and red) compared with the highest concentration of SYBR Green I (purple and yellow) that did not exhibit PCR inhibition, leading to improved melt curve & HRM performance.

Specifications

Protocol

Store Chai Green at -20 ºC and protect from light. Chai Green is provided as a 20X solution in water. Use the dye at a 1X concentration in the qPCR reaction. Add 1.25 μL 20X Chai Green per 25 μL reaction. Use with a thermocycler capable of measuring SYBR Green I or FAM fluorescence during the extension phase of qPCR. The 1X working concentration may also be used for melt curve analysis and High Resolution Melt (HRM).

License

To promote collaboration in science, Chai Green Dye 20X is provided under an open license:

• It may be used freely for any application, including commercial and diagnostic use
• No license agreements or royalty payments required for OEM use
• May be incorporated into your own test kits without restriction

Resources