RealHelix™ qRT-PCR Kit [Probe] is a convenient and reliable one-step quantitative RT-PCR kit. Both cDNA synthesis and PCR reactions are performed in a single tube using sequence-specific primers corresponding to the target RNAs from total RNA or mRNA. The Enzyme Mix in this kit is an optimized blend of HelixCript™ Thermo Reverse Transcriptase, RNase inhibitor and a specially designed automatic hot-start version of PCR enzyme, and ensures reliable results in terms of sensitivity, accuracy and specificity. The 2x Buffer Mix contains all of the required components including optimized buffer components, Mg²⁺, dNTPs. The separately supplied ROX dye could be used as a passive reference dye in real-time PCR instruments that are compatible with the evaluation of the ROX signal.

Figure 1. Multiplex (four targets) qRT-PCR with RealHelix™ qRT-PCR Kit [Probe].

Total RNAs from human whole blood were analyzed using four different sets of target-specific primers and probes. The probes were labeled with FAM, HEX, Texas Red, or Cy5. The multiplex reaction was performed using the RealHelix™ qRT-PCR Kit [Probe] on a real-time PCR instrument, the CFX96 (Bio-Rad, USA).

Figure 2. Comparison of RealHelix™ qRT-PCR Kit [Probe] and qRT-PCR Kit [Probe type] (the previous version).

10-fold serial dilutions (10, 1, 0.1, and 0.01 ng) of the human total RNA from whole blood were analyzed using a human globin gene specific primer set. RealHelix™ qRT-PCR Kit [Probe] shows earlier Ct and more reliable amplifications than the previous version. Reactions were run on a real-time PCR instrument, CFX96 (Bio-Rad, USA).

Storage

• Store below -20 ℃
• ROX dye should be stored in a dark place

Shelf life

• 12 months