1-Sec qRT-PCR Premix [Probe] is designed to perform rapid real-time analysis of RNA samples using fluorescent probe based detection. This kit provides 2x premix and separates ROX reference dye. The convenient 2x concentrated premix contains Ab+Taq polymerase, Thermo reverse transcriptase, RNase inhibitor, dNTPs, buffers, Mg2+, and stabilizing agent. The premix can also be used in combination with ROX reference dye in PCR instruments that are compatible with the evaluation of the ROX signal. The outstanding fast real-time assay within 40-60 min combined with high specificity and sensitivity is achieved with a unique buffer system and balanced enzyme mix. The w/UDG version additionally contains a heat-labile UDG and dUTP in the premix. The applied UDG system prevents carryover contamination.
Application
- Fast real-time PCR analysis with labeled-probes
- Multiple target detection and quantification
- SNP analysis and Genotyping
- Microbial & viral pathogen detection
- Molecular diagnosis
Figure 1. Comparison of amplification plots of a general qRT-PCR enzyme (RealHelix™ qRT-PCR kit) and 1-sec qRT-PCR Premix [Probe] (= RealHelix™ 1-sec qRT-PCR kit [Probe]) using either a standard cycling protocol or a fast cycling protocol. Multiplex Probe qRT-PCR reactions were done with the purified and mixed RNA of type A virus and type B virus (tenfold serial dilutions over range of 1 fg - 1 ng per reaction). Each 20μl reactions were operated on CFX96 real-time PCR detection system (BioRad).
Properties: Real-time RT-PCR, Probe type, Hot-start, Fast PCR
Heat activation: +95°C for 2 minutes after the reverse transcription reaction.
Activity test: Performance test in PCR using Bio-Rad CFX-96. (Human total RNA, β-globin and GAPDH fragment): Corresponds to specification