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RealHelix™ qRT-PCR kit

● Convenient real-time RT-PCR in one-tube reaction.
● Ready-to-use buffer mix.
● Reliable quantification of target RNA.
● High specific amplification using automatic hot-start enzyme.
● ROX passive reference dye provided.
● Contains a strong fluorescent dye.


Description

RealHelix™ qRT-PCR kit 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 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 the required components including optimized buffer components, Mg2+, dNTPs, and RNase inhibitor. Two-types of qRT-PCR kits are available. qRT-PCR [Intercalator type] is dedicated for SYBR Green detection, and qRT-PCR [Probe type] is for the detection using sequence-specific probes. ROX passive reference dye is also provided with separated tube for normalization of the fluorescence signal.


Application
◎ Detection of target gene transcript from RNA in one-tube format
◎ Absolute and relative quantification analysis of RNA transcription level
◎ Molecular Diagnostics for RNA virus

Contents

◎ Enzyme Mix
◎ 2x Buffer Mix with Evagreen fluorescent dye
◎ ROX Passive Reference Dye


Quality Control

The activity for cDNA synthesis and quantitative PCR of target transcript using RealHelix™ qRT-PCR Kit was evaluated by real-time PCR using the 10-fold serial-diluted total RNA isolated from human blood and beta-globin gene-specific primer set with ABI 7900HT fast-real-time PCR system(Applied Biosystems, USA) and Rotor-Gene Q instrument (QIAGEN, Germany).


Data

Figure 1. Comparison of one-step qRT-PCR using qRT-PCR Kit [Intercalator type] and Rotor-Gene SYBR RT-PCR Kit. Total RNAs (10-fold diluted RNAs from 10 to 0.1 ng) isolated from human blood were analyzed using the RealHelix™ qRT-PCR Kit or Rotor-Gene SYBR Green RT-PCR Kit, and a primer set specific Human β-globin gene gene. Triplicated reactions were run on a real-time PCR instrument, Rotor-Gene Q (QIAGEN, Germany). Melting-curve analysis revealed the high specificity of qRT-PCR. The accuracy of qRT-PCR is demonstrated by R2 value = 0.996 (RealHelix™ qRT-PCR, NanoHelix) and R2 value = 0.991 (Rotor-Gen SYBR Green RT-PCR Kit).

 

Figure 2. Sensitive detection using qRT-PCR Kit [Intercalator type].Total RNAs (10-fold diluted RNAs from 10 ng to 10 pg) isolated from human blood were analyzed using the qRT-PCR Kit and primer set specific human β-globin gene. Quadrupled reaction were run on a real-time PCR instrument, Rotor-Gene Q (QIAGEN, Germany).

 

Figure 3. Real-time PCR for diagnostics of HAV using qRT-PCR Kit [Probe type]. Total RNA isolated from HAV particles using PureHelix™ Total RNA purification Kit. Real-time detections were performed according to the manual of qRT-PCR kit. NTC : No Template Control.
Cat.No. Product Size
 QRT-S100 RealHelix™ qRT-PCR kit [Intercalator type] (ROX Solution provided) 100 rxns 
 QRT-S500 RealHelix™ qRT-PCR kit [Intercalator type] (ROX Solution provided) 500 rxns 
 QRT-P100 RealHelix™ qRT-PCR kit [Probe type] (ROX Solution provided) 100 rxns 
 QRT-P500 RealHelix™ qRT-PCR kit [Probe type] (ROX Solution provided) 500 rxns