Premier 2x qPCR Mix [Green] is designed to perform a rapid, highly specific, and sensitive real-time quantification of target DNA. Specially designed buffer and antibody-mediated hot start enzyme considerably reduce the primer dimer formations and ensure producing reliable data. The convenient 2x concentrated premix contains Ab+Taq polymerase, dNTPs, buffers, Mg2+, a green fluorescent dye, and stabilizing agent. Carryover contaminated PCR products could be removed by applying the UDG system.
Application
- Real-time quantification of target DNA
- Automated high throughput real-time PCR
- Gene expression analysis of cDNA
- SNP analysis and Genotyping
- Microbial & viral pathogen detection
- Molecular diagnosis
Figure 1. Sensitive detection using Premier 2X qPCR Mix [Green] (= RealHelixTM Premier qPCR Mix [Green]). 10-fold serial-diluted genomic DNA as the standard sample were analyzed using the Premier 2X qPCR Mix [Green] for human glucose-6-phosphate dehydrogenase gene-specific primer set. The accuracy of real time PCR using the Premier 2X qPCR Mix [Green] is demonstrated by R² value=0.99887. NTC: Negative control.
Figure 2. Comparison of Real-time PCR using Premier 2X qPCR Mix [Green] (= RealHelix™ Premier qPCR Mix [Green]) 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 (Premier qPCR Mix), 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 Premier 2X qPCR Mix [Green] (= RealHelix™ Premier qPCR Mix [Green]) 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 Mix [Green]), 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 Premier qPCR Mix. 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 Premier 2X qPCR Mix [Green] (= RealHelix™ Premier qPCR Mix [Green]). Premier 2X qPCR Mix [Green] 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.
Properties: Real-time PCR, Intercalator type, Hot-start
Heat activation: +95°C for 2 minutes
Activity test: Performance test in PCR using Bio-Rad CFX-96 (Human genomic DNA, G6PDH fragment): Corresponds to specification
Stability: 12 months at -20°C
NanoHelix的 Multiplex PCR 2x Premix是设置复杂多重PCR的最佳选择，是一种预混合溶液，其中包含HelixAmp™ Hot-Taq聚合酶，dNTP和优化的缓冲液，浓度为2x。Multiplex PCR 2x Premix专为热启动PCR设计，可实现最高的特异性并最大程度地减少反应混合物中引物之间的干扰。残留污染的PCR产物可以通过使用UDG系统去除。 应用 多重PCR（常规）等位基因特异性PCRSNP分析和基因分型
