The probe-based qPCR experiment is a type of real-time fluorescence quantitative PCR. Based on its high specificity, high sensitivity, real-time and quantitative detection advantages, it has been used in pathogen detection, Gene expression analysis It has been widely used in many biological and medical research fields such as genotype identification, copy number variation analysis, food safety testing, environmental testing, and forensic testing.
The development of detection and diagnostic reagent products based on probe-based qPCR technology integrates upstream molecular enzyme raw materials, sample pre-treatment, detection reagent reaction system optimization, Real-time fluorescence quantitative PCR instrument There are multiple links including research and development, registration and declaration (which varies according to product projects), etc.
In many aspects, Upstream molecular enzyme raw materials It is the starting point and key point of the entire product development process. Many companies either develop molecular enzymes from scratch, purchase enzyme raw materials separately, directly purchase qPCR mixes optimized by raw material manufacturers, or outsource the entire reagent process.
The market is so large that each company has its own corresponding strategy. Regardless of the method, the goal is to provide a stable product line that can truly solve problems and has high cost-effectiveness.
qPCR Mix usually contains other components required for the reaction in addition to template DNA, primers and probes. Among them, the molecular enzyme raw materials and buffer system play an important role. How to select or optimize the appropriate qPCR Mix is the key to the success of probe-based qPCR experiments. Based on years of work experience, the R&D team of Qihengxing Bio has summarized the factors that need to be considered in qPCR Mix screening and optimization and the screening steps from different levels.
| Project Name | Project Content |
| Purpose of the experiment | Determine the target gene type to be detected in the experiment, such as gene expression analysis, pathogen detection, target gene mutation analysis, etc. Select a qPCR mix that is compatible with the target sequence. |
| Amplification efficiency requirements | Try to choose a highly efficient qPCR Mix to improve the sensitivity and specificity of the experiment. |
| Experimental throughput | Select the desired qPCR Mix based on singleplex or multiplex detection requirements. |
| Stability and convenience | Choosing a premix can simplify experimental operations and reduce errors. At the same time, the stability and convenient storage conditions of the premix are also factors that need to be considered; choose a qPCR Mix with high thermal stability. |
| Anti-pollution | Add anti-contamination components (such as UNG, etc.) to the qPCR Mix to reduce false positive results. |
| Cost-effectiveness | Consider your experimental budget and choose a cost-effective qPCR Mix. |
| Brands and Suppliers | Choose brands and suppliers with good reputation and high service quality; consider the supplier's technical support and product after-sales service. |
| User Reviews | Refer to other researchers' usage experiences and reviews. |
Factors to Consider in Probe-Based qPCR Mix Screening and Optimization
Probe-based qPCR Mix Screening Steps
| Application Scenario | Product Recommendations |
| SNP genotyping test | The first choice is FS-Q2001, followed by FS-Q6001. |
| Pathogens, viruses, nucleic acid residues, low copy gene detection, etc. |
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| Methylation testing | FS-Q6001 is recommended. |
| Bold samples (testing samples containing inhibitors) | FS-Q6001 is recommended. |
| Melting curve method (probe method qPCR mix) | FS-Q6001 is recommended. |
| Any of the above 1-5 application scenarios | Considering the diversity of probe-based qPCR reaction systems, you can also test FS-Q2001, FS-Q3001, FS-Q3010, and FS-Q6001 simultaneously and select the most suitable one. |
| Unable to meet customer needs | The company's four existing products all provide CDMO customized services |
Probe-Based qPCR Mix Selection Guide
Probe-Based qPCR Mix Product List
