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RealHelix™ Premier qPCR Kit [SYBR Green with ROX]

 

● ROX passive dye pre-mixed
● Rapid quantification of target DNA
● High specific amplification using antibody mediated hot-start enzyme


Description

RealHelix™ Premier qPCR Kit [SYBR Green with ROX] is designed to perform a rapid real-time quantification of target DNA or first strand cDNA synthesized from RNA molecule. The 2x premix contains SYBR Green I dye, ROX passive dye, antibody-mediated hot-start DNA polymerase, dNTPs, buffer, MgCl2 stabilizers, and enhancers.


Application
◎ Quantification of target DNA sample by real time PCR

Contents

2x Premier qPCR PreMix


Quality Control

RealHelix™Premier qPCR Kit [SYBR Green with ROX] was evaluated by real-time PCR using the 10-fold serial-diluted human genomic DNA and a set of human gene-specific primer with ABI7500 real-time PCR system(Applied Biosystems, USA), Rotor-Gene Q instrument(QIAGEN, Germany), and Bio-Rad CFX96(Bio-Rad, USA).

 

 

Data




Figure 1. Sensitive detection using RealHelix™ Premier qPCR Kit [SYBR Green with low ROX]. 10-fold serial-diluted genomic DNA as the standard sample were analyzed using the RealHelix™ Premier qPCR Kit for human glucose-6-phosphate dehydrogenase gene-specific primer set. The accuracy of real time PCR using the RealHelix™ Premier qPCR Kit is demonstrated by R2 value=0.99887. NTC: Negative control.

 



Figure 2. Comparison of Real-time PCR using RealHelix™ Premier qPCR Kit [SYBR Green with low ROX] iTaq Universal SYBR green supermix (company B) and TOPreal qPCR 2x premix (company E). 10-fold serial dilutions of the human genomic DNA (red: 50 ng/rxn, green: 5 ng/rxn, blue: 0.5 ng/rxn) from whole blood were analyzed using human HPRT-1 gene-specific primer set. The accuracy of real-time PCR is demonstrated by R2 value=0.99482 (RealHelix™ Premier qPCR Kit), R2 value=0.98909 (iTaq Universal SYBR green supermix) and R2 value=0.99233 (TOPreal qPCR 2x premix). Reactions were run on a real-time PCR instrument, Rotor-GENE Q (QIAGEN, Germany).

 

Figure 3. Comparison of Real-time PCR using RealHelix™ Premier qPCR Kit [SYBR Green with low ROX] iTaq Universal SYBR green supermix (company B) and TOPreal qPCR 2x premix (company E). 10-fold serial dilutions of the human genomic DNA (red: 50 ng/rxn, green: 5 ng/rxn, blue: 0.5 ng/rxn) from whole blood were analyzed using human HPRT-1 gene-specific primer set. The accuracy of real-time PCR is demonstrated by R2 value=0.999 (RealHelix™ Premier qPCR Kit), R2 value=0.999 (iTaq Universal SYBR green supermix) and R2 value=1.000 (TOPreal qPCR 2x premix). Melting curve analysis revealed the high specificity of RealHelix™ Premier qPCR Kit. Reactions were run on a real-time PCR instrument, CFX-96 (Bio-Rad, USA). NTC: Negative control.

 

Figure 4. Amplification of target DNA with various G+C contents using RealHelix™ Premier qPCR Kit [SYBR Green with low ROX]. RealHelix™ Premier qPCR Kit [SYBR Green with low ROX] was evaluated by real-time PCR using 10-fold serial diluted human genomic DNA (red: 50 ng/rxn, green: 5 ng/rxn, blue: 0.5 ng/rxn) with Rotor-Gene Q instrument (QIAGEN, Germany), CFX-96 instrument (Bio-Rad, USA) and ABI7500 real-time PCR system (Applied Biosystems, USA). NTC: Negative control.

 

 

Cat.No. Product Size
 PQL-S200 RealHelix™ Premier qPCR Kit [SYBR Green with low ROX] 200 rxns 
 PQL-S500 RealHelix™ Premier qPCR Kit [SYBR Green with low ROX] 500 rxns 
 PQH-S200 RealHelix™ Premier qPCR Kit [SYBR Green with high ROX] 200 rxns 
 PQH-S500 RealHelix™ Premier qPCR Kit [SYBR Green with high ROX] 500 rxns