Introduction
Digital PCR (also known as single-molecule PCR) generally consists of two parts, PCR amplification and fluorescence signal analysis. In the PCR amplification phase, unlike conventional techniques, digital PCR generally requires diluting the sample to a single molecule level and distributing it evenly over tens to tens of thousands of units. Unlike the method in which qPCR performs real-time fluorescence measurement on each cycle, the digital PCR technique collects the fluorescence signal of each reaction unit after the amplification is completed. Finally, the original concentration or content of the sample is calculated by direct counting or Poisson distribution formula.
Functional
1. The fluid control system for sample preparation is simple, one chip can simultaneously prepare droplets of 8 samples and control independently between channels;
2. Innovative droplet preparation of the oil phase, compared with the commonly used fluorinated oil on the market, can avoid the generation of bubbles during the formation of droplets;
3. The sample chip is self-developed, and the sample size is uniform. The single layer is stably tiled in the chip to avoid cross-contamination and easy to detect and analyze.
4. Multi-channel real-time fluorescence imaging analysis, direct one-time dynamic scanning, no sample loss;
5. The reader can realize the identification analysis of liquid chips and solid chips;
6. The whole system has a high throughput and detects a throughput of 32 gene chips;
Application
The application of digital PCR technology in genomics research
1. Research on genomic expression and regulation:
This field requires the recognition of all genomic expression products mRNA and protein and their interaction at the whole cell level; digital PCR technology is well suited for high-precision quantification of gene expression products, especially microRNAs, and can achieve single cell and single molecular weight levels. Quantitative detection and analysis of nucleic acids.
2. Identification and identification of human genetic information:
To extract genomic function information, identifying and identifying gene sequences is essential for basic work. Digital PCR technology can verify and confirm the sequence of the gene obtained by sequencing, especially in the case that the original sample is difficult to obtain or the original content is extremely low, because digital PCR technology is good at identifying weak signals from the background signal.
3. Extraction and identification of gene function information:
Specifically, it includes systematic identification of human gene mutants, mapping of gene expression profiles, and identification of "gene alteration-function changes"; digital PCR technology can achieve gene expression profiling and highly sensitive detection of gene mutants.
4. Genetic polymorphism analysis:
Although the genome sequence obtained by the Human Genome Project is representative, each person's genome is not exactly the same. The most common polymorphism is single nucleotide polymorphisms (SNPs), and digital PCR technology passes the reaction. The unit is discretized and has a small influence on the amplification efficiency of the polymerase chain reaction (PCR), and can achieve a SNP analysis with a sensitivity of one hundred thousandth.
The application of digital PCR technology in translational medicine and precision medicine
1. Individualized diagnosis and treatment of cancer
2. Ultra-early diagnosis of disease - "liquid biopsy"
3. Non-invasive prenatal examination
4. Pathogenic microorganism detection and other disease prevention and control
bio-equip.cn
Pilot Gene Technology (Hangzhou) Co., Ltd.is a high-tech enterprise company specializing in the development, production, sales and service of life science instruments and reagents. The company uses PCR (Digital PCR, dPCR) technology to build a high-tech diagnostic technology platform for independent innovation, independently develops the production of digital PCR products, and is committed to the development of diagnostic reagents in the three major clinical fields of hereditary diseases, accurate diagnosis and treatment of tumor diseases and infectious diseases.
The company has a high level of multi-disciplinary research and development team and high quality senior sales team, and established the international top R & D center and GMP production base, won the "Eagle plan" enterprises and national high-tech enterprises to identify, and has nearly thirty independent intellectual property invention patents.
Through the implementation of modern management system and mutual benefit and win-win cooperation mechanism, the East China gene has continuously opened up the international research and development team, built a better production base and a rigorous quality system, striving for the continuous breakthrough and innovation of core technology, and striving to become the top upstream supplier in the field of domestic liquid inspection. Medical institutions, governments and people win a lot in order to make greater contributions to the cause of human health.
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