2x qPCR Premix [Green] is designed to perform a rapid real-time quantification of target DNA. The convenient 2x concentrated premix contains hot-start PCR enzyme, dNTPs, buffers, Mg2+, a fluorescent green dye, and stabilizing agent. The hot-start PCR enzyme provides high specific amplification of target DNA and minimizes the non-specific reactions and production of primer dimers. Carryover contaminated PCR products could be removed by applying the UDG system.
Application
- Real-time quantification of target DNA
- Automated high throughput real-time PCR
- Gene expression analysis of cDNA
- SNP analysis and Genotyping
- Microbial & viral pathogen detection
- Molecular diagnosis
The accuracy of real-time PCR using 2x qPCR Premix [Green] (= RealHelix™ qPCR Kit [Green])
Human genomic DNAs (10, 1, 0.1, 0.01ng) isolated from whole blood were analyzed using alpha 1-Antitrypsin(AAT) DNA-specific primer set and an intercalating dye. The accuracy of real-time PCR is demonstrated by E=107.7%, R² value = 0.999. 20 µl reactions were operated on a CFX96 real-time PCR detection system (BioRad).
Comparison of real-time PCR using 2x qPCR Premix [Green] (= RealHelix™ qPCR Kit [Green]) and the previous version (RealHelix™ qPCR Kit [Intercalator type])
Human genomic DNAs (100, 10, 1, 0.1ng) isolated from whole blood were analyzed using beta-globin gene-specific primer set and an intercalating dye.
Each 20 µl reactions were operated on a CFX96 real-time PCR detection system (BioRad).
2x qPCR Premix [Green] shows earlier Ct and more efficient amplification.
Properties: Real-time PCR, Intercalator type, Hot-start
Heat activation: +95°C for 15 minutes
Activity test: Performance test in PCR using Bio-Rad CFX-96 (Human genomic DNA, Actin fragment): Corresponds to specification
Stability: 12 months at -20°C