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Bst DNA聚合酶，大片段是嗜热脂肪芽孢杆菌 DNA聚合酶蛋白的一部分，具有5'→3'聚合酶活性，但缺乏5'→3'核酸外切酶活性。Bst DNA聚合酶，大片段作为重组体从大肠杆菌菌株中纯化。 应用 回路介导的等温扩增（LAMP）DNA链置换扩增全基因组扩增
