The RealHelix™ qRT-PCR kit [Probe] is a convenient and reliable one-step quantitative RT-PCR kit using dual-labeled fluorescence probe. The Enzyme Mix in this kit is an optimized blend of Reverse Transcriptase, hot-start PCR enzyme, and RNase inhibitor protein, which ensures reliable results regarding 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^TM qRT-PCR[Probe] Kit. 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 done using the RealHelix^TM qRT-PCR Kit [Probe] on a real-time PCR instrument, Bio-Rad CFX96. 

Figure 2. Comparison of RealHelix^TM 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 human Globin gene specific primer set. RealHelix^TM qRT-PCR Kit [Probe] shows earlier Ct and more reliable amplifications than the previous version. Reactions were run on a real-time PCR instrument, CFX-96 (Bio-Rad, USA).

Contents 

• QRT Enzyme Mix [Probe]
• QRT 2x Buffer Mix [Probe]
• ROX Dye (25μM) 

Application 

• Probe-based real-time quantification of target RNA
• Multiple detections of target RNAs
• Gene expression analysis 
• Viral RNA detection.
• Molecular diagnosis 

Quality control

RealHelix™ qRT-PCR Kit [Probe] was evaluated by real-time RT-PCR using the 10-fold serial-diluted human total RNA and a set of human gene-specific primer and dual-labeled probe with ABI 7500 real-time PCR system (Applied Biosystems, USA) and CFX96™ real-time PCR detection system (Bio-Rad, USA).