● Fast : 30 mins for 40 cycles (1-sec denature, 1-sec anneal / extension)
● Multiplex : Up to 5-plex probe qPCR in a reaction
● High sensitivity : Reliable detection as low as 10 copies of target.
● Robust hot-start enzyme : Novel Antibody inhibited Taq polymerase
RealHelix™ 1-sec qPCR Kit [Probe type] is designed to perform a rapid real-time quantification of target DNA or 1st-strand cDNA using dual-labeled probe. The 2x premix contains enzymes and buffers which are designed for higher processivity and speed, offering significantly faster extension rates than other qPCR products. The kit consists of convenient 2x concentrated premix solution and ROX passive dye. The 2x premix contains antibody-mediated hot-start PCR enzyme, 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. Included ROX dye could be used as a passive reference dye.
|◎ Quantification of target DNA sample by real-time PCR |
◎ 2x 1-sec qPCR Premix
◎ ROX Passive Reference Dye
RealHelix™ 1-sec qPCR Kit was evaluated by real-time PCR using the 10-fold serial-diluted human genomic DNA 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).
|Figure 1. Comparison of amplification plots of a general qPCR enzyme (RealHelix™ qPCR kit, probe, type) and 1-sec qPCR kit using either a standard cycling protocol or a fast cycling protocol. Multiplex Probe qPCR reactions were done with a purified human genomic DNA over range of 0.01 ng to 10 ng per reaction. Each 20μl reactions were operated on CFX96 real-time PCR detection system (BioRad).|
|Figure 2. GloB gene fragment was amplified from 10-fold serial dilutions of human genomic DNA (10 ng - 0.1 ng/20μl reaction) using 1-sec qPCR Kit or other competitor's fast probe qPCR kits; Competitor A (Probe Fast qPCR kit) and Competitor B (Universal Probe supermix) in fast condition. Fluorescence data were obtained in a multiplex (3-plex) qPCR reactions.|