Chai Green qPCR Master Mix
- Produce excellent melt curves & HRM results with the highly fluorescent Chai Green dye
- Overcomes inhibition in crude extracts and environmental samples
- Universal mix runs fast protocols, amplifies GC-rich targets, and incorporates aptamer-based hot start and dUTP carry-over prevention systems
- License-free for commercial or diagnostic use
Chai Green qPCR Master Mix is a 2X Taq mix for dye-based Real-Time PCR, melt curve analysis, and High Resolution Melt (HRM). It quantitatively amplifies singleplex and duplex qPCR targets at high efficiency from purified samples and crude extracts.
Better Melt Curves & HRM
As Chai Green is less inhibitory to PCR than SYBR Green, it can be used at significantly higher concentrations. The resulting increased fluorescence improves the ability to differentiate small differences via melt and HRM analysis.
Amplifies Everything Without Inhibition
Chai Green qPCR Master Mix has been validated against genomic, plasmid, and cDNA targets from a wide range of organisms. The mix supports both standard and fast protocols, and amplifies GC-rich targets up to 70%.
Thanks to numerous enhancers which bind & inactivate inhibitors, the mix resists inhibition in crude extracts from:
- Food & beverages
- Plant & animal tissue
- Environmental air & water samples
Simplify your PCR
Set up reactions with ease at room temperature: an aptamer-based hotstart system prevents amplification from occurring below 50 ºC, while a visible loading dye helps avoid pipetting errors. When activated by the addition of UNG, a dUTP carry-over prevention system prevents false positives from amplicon contamination.
Exceptional Linearity & Dynamic Range
Dynamic range of 1010 is demonstrated while maintaining excellent efficiency and linearity using Chai Green qPCR Master Mix to amplify a 157 bp gene fragment on an Open qPCR instrument.
Clear Peaks and Tm Multiplexing
Reactions containing mActin, mActin + Lambda, and Lambda + mGAPDH targets were amplified using Chai Green qPCR Master Mix on an Open qPCR instrument. Peaks can clearly be identified and differentiated, even when multiple targets are present.
A class 4 beta globin SNP (rs7936221) was genotyped using Chai Green qPCR Master Mix. Wild type, homozygous, and heterozygous variants were easily differentiated via HRM, despite the class 4 SNP’s Tm difference of only 0.2 ºC.
Fast & Quantitative
A fast 19 minute protocol was run on an Open qPCR instrument, obtaining excellent linearity & efficiency. Protocol: Initial denaturing 30 s @ 95 ºC, Cycle 30x: 3 s @ 95 ºC, 14 s @ 60 ºC, 5 C ºC/s ramp times.
Fast & Effective Hot Start
A duplex lambda phage (72 bp) and salmon sperm (77 bp, 67% GC content) assay was run with and without Chai Green qPCR Master Mix’s hot start aptamer. When run with 5 ng/μL of template DNA, both targets amplified regardless of hot start ability. When run with the more dilute 0.0005 ng/μL, the salmon sperm target failed to amplify without hot start due to non-specific products forming, while the hot start aptamer prevented these non-specific products, allowing both targets to amplify.
|Hot Start Mechanism||Aptamer|
|GC Content||40 – 70%|
|Multiplex Targets||Up to 2|
|dUTP Carryover Prevention||Yes†|
|Visible Loading Dye||Yes|
|Exonuclease Activity||5' → 3'|
|Supported Templates||Genomic DNA, cDNA, Plasmid DNA|
|Recommended Reaction Volume||10 – 50 µL|
† user must add UNG to enable
Shipping, Storage, and Stability
|High Temperature Stability||2 weeks @ 25 ºC|
|Freeze/Thaw Cycles||Up to 20|
|Storage Conditions||-20 ºC long-term, or 4 ºC for 6 months. Protect from light|
The mix ships with an optional ROX reference dye, allowing it to be used with high, low, and no-ROX qPCR instruments.
What is a PCR Master Mix?A PCR master mix is a premixed solution that contains most of the components necessary to run a PCR assay. The mix contains Taq DNA polymerase, dNTPs, MgCl2, as well as enhancers and stabilizers in a buffer that is optimized for DNA amplification by PCR. Read more...
To promote collaboration in science, Chai Green qPCR Master Mix 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