RealHelix™ qRT-PCR kit [Green] 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 the sample. 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, and a green fluorescent dye. 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.  

Comparison of RealHelix™ qRT-PCR Kit [Green] and qRT-PCR Kit [Intercalator type] (the previous version). 10-fold serial dilutions of the human total RNA (1, 0.1, and 0.01 ng) from whole blood were analyzed using human beta actin gene-specific primer set. RealHelix™ qRT-PCR Kit [Green] shows more reliable amplifications than our previous version. Reactions were run on a real-time PCR instrument, CFX-96 (Bio-Rad, USA). 

Contents 

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

Application 

• Real-time quantification of target RNA
• Automated high throughput one-step real-time RT-PCR
• Gene expression analysis
• Viral RNA detection
• Molecular diagnosis 

Quality control

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