Research and Application of Fluorescence Quantitative PCR
Principles of real-time PCR
Fluorescence quantitative PCR (Real-time PCR) refers to the addition of fluorescent groups to the PCR amplification reaction system, through the real-time detection of the fluorescent signal of each cycle product in the amplification reaction, and finally the quantitative analysis of the unknown template through the standard curve Methods.
Application of Fluorescence Quantitative PCR
Molecular biology research
1. Nucleic acid quantitative analysis: quantitative and qualitative analysis of infectious diseases, detection of pathogenic microorganisms or virus content, such as the recent epidemic of H1N1 influenza, detection of gene copy number of genetically modified animals and plants, detection of RNAi gene inactivation rate, etc.
2. Gene expression difference analysis: compare the expression differences of specific genes between samples after different treatments (such as drug treatment, physical treatment, chemical treatment, etc.), the expression differences of specific genes in different phases, and the confirmation of the cDNA chip or difference display results
3. SNP detection: The detection of single nucleotide polymorphisms is of great significance for studying the susceptibility of individuals to different diseases or the different responses of individuals to specific drugs. Due to the ingenuity of the molecular beacon structure, once the sequence information of the SNP is It is known that high-throughput SNP detection using this technology will become simple and accurate.
4. Methylation detection: Methylation is related to many human diseases, especially cancer. Laird reported a technology called Methylight, which processes DNA before amplification so that unmethylated cytosine becomes Uracil, while methylated cytosine is not affected. Using specific primers and Taqman probes to distinguish between methylated and unmethylated DNA is not only convenient but also more sensitive.
Medical Research
1. Prenatal diagnosis: People are still unable to treat genetic diseases caused by genetic material changes. So far, only prenatal monitoring can be used to reduce the birth of sick babies to prevent the occurrence of various genetic diseases. For example, in order to reduce the birth of children with X-linked genetic diseases, separating fetal DNA from the peripheral blood of pregnant women and detecting the Y sex-determining region genes by real-time fluorescent quantitative PCR is a non-invasive method that is easily accepted by pregnant women.
2. Pathogen detection: fluorescent quantitative PCR detection technology can detect Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum, human papilloma virus, herpes simplex virus, human immunodeficiency virus, hepatitis virus, influenza virus, Mycobacterium tuberculosis, Epstein-Barr virus And cytomegalovirus and other pathogens for quantitative determination. Compared with traditional detection methods, it has the advantages of high sensitivity, less sampling, quickness and simplicity.
3. Evaluation of drug efficacy: Quantitative analysis of hepatitis B virus (HBV) and hepatitis C virus (HCV) shows that the amount of virus is related to the efficacy of certain drugs. HCV is expressed at high levels and is not sensitive to interferon treatment, while HCV is low in titers and interferon is sensitive; during lamivudine treatment, the serum level of HBV-DNA has declined, and then if it rises again or exceeds the previous level , It indicates that the virus has mutated.
4. Tumor gene detection: Although the mechanism of tumor pathogenesis is not yet clear, it is widely accepted that mutations in related genes are the root cause of carcinogenic transformation. Increased expression and mutation of oncogenes can appear in the early stages of many tumors. Real-time fluorescent quantitative PCR can not only effectively detect gene mutations, but also accurately detect the expression of oncogenes. At present, this method has been used to detect the expression of multiple genes such as telomerase hTERT gene, chronic myelogenous leukemia WT1 gene, tumor ER gene, prostate cancer PSM gene, tumor-related viral genes and so on.
Yongyue Medical's fluorescent quantitative PCR plates and fluorescent quantitative PCR tubes can meet the different needs of our customers. The products are produced in a 100,000-level purification workshop. The quality is safe and reliable, and the quality is excellent. It is recognized by the majority of users and helps scientific experimental research and Innovation.
Features of fluorescent quantitative PCR tube:
Medical grade PP raw material, in line with USP VI grade, to ensure low liquid loss;
The product is easy to close, smooth to open, and easy to operate;
Ultra-thin and uniform tube wall ensures the best heat conduction effect and good light transmittance;
Produced in a 100,000-level purification workshop, free of DNase, RNase and endotoxin pollution;
The product has excellent properties such as optics, air tightness, and stable deformation.
Product specifications: The capacity is divided into 0.1ml and 0.2ml; the categories are single tube, 8 continuous rows of tubes/caps.
