RealHelix™ qPCR Kit [Green] is designed to perform a rapid real-time quantification of target DNA using a double strand DNA-binding green fluorescent dye. The convenient 2x premix contains hot-start PCR enzyme, green fluorescent dye, dNTPs, buffers, Mg2+, and stabilizing agent. The hot-start PCR enzyme provides high specific amplification of target DNA and minimizes the side products such as primer dimers.
The accuracy of real-time PCR using 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 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). RealHelix™ qPCR Kit [Green] shows earlier Ct and more efficient amplification.
Contents
- 2x Premix [Green]
- ROX Dye (25μM)
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
Quality control
RealHelix™ qPCR Kit [Green] was evaluated by real-time PCR using the 10-fold serial-diluted human genomic DNA and a set of human gene-specific primer with ABI 7500 real-time PCR system (Applied Biosystems), CFX96 (BioRad), and Rotor-Gene Q instrument (QIAGEN).