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Application and development of PCR cloning

Update time : 2020-08-11

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Application and development of PCR cloning

The polymerase chain (PCR) reaction technology has many advantages such as strong specificity, high sensitivity, rapidity and simplicity, and can amplify the desired target gene to tens of thousands of times in a short time. 

Today, with the rapid development of life sciences, PCR cloning technology has become a routine technology in molecular biology. PCR cloning technology has been widely used in biological research, medicine, virus detection, food industry and so on.

PCR technology applied to gene cloning

The use of PCR technology for gene cloning and subcloning has played a huge role in cell biology research. 

PCR cloning technology can amplify a single copy of a gene several million times, and the generated specific DNA fragments can be micrograms (pg), omitting the DNA digestion required to clone a specific gene fragment from genomic DNA. 

The tedious experimental process of ligation to vector DNA, transformation, establishment of DNA library, gene screening and determination, and subcloning.

PCR cloning technology applied to DNA recombination

In molecular biology research, specific genes or DNA fragments from different sources can be inserted into viruses, plasmids or other vectors in vitro by PCR cloning technology to construct recombinant DNA molecules, and then introduce them into recipient cells for amplification and reproduction. 

After screening, the transformant cells containing the target gene are amplified and extracted to obtain a large amount of DNA. 

Recombinant DNA technology can be applied to research in the human genome project, valuable protein expression, gene diagnosis and treatment, genetically modified plants and animals, and other fields.

Application of PCR cloning technology in gene quantitative detection

The PCR cloning technology can quantitatively monitor the copy number of the target gene in the specimen, and place the target gene and a single copy of the reference gene in the same test tube for PCR cloning. 

Observe the band intensity after electrophoresis separation, or label a radionuclide at the 5'end of the primer, and detect the gene copy number by detecting the radioactivity intensity of the two bands. 

Using PCR cloning technology can also complete the quantitative detection of mRNA. 

First, advance the total RNA, add primers complementary to regions 3 and 7 of mRNA, synthesize cDNA under the action of reverse transcriptase, and then refer to the steps of DNA quantitative detection.

The application of PCR cloning technology can accurately quantify tuRNA, and even 1TIRNA that could not be found by Northern blot hybridization can be detected.

The change of endogenous genes and the invasion of foreign genes are also a great threat to human diseases. However, regardless of whether the cause of the disease is caused by genetic changes, as long as the pathogen exists, the nucleic acid can be found. 

As a result, PCR cloning technology and its related technologies have been developed so far, and they can be fully applied to the detection and diagnosis of infectious disease pathogens, tumor-related gene detection, early diagnosis of genetic diseases, bone marrow transplantation HLA-D site matching and chemical analysis and forensic medicine Research in other fields.

Generalbiosystems' patented cloning technology can help customers clone the target gene to any designated position of the desired vector without relying on the vector restriction site, satisfying customers' various cloning ideas, and greatly saves compared to ordinary cloning Time. 

For the same vector cloning of different gene fragments, we can also efficiently complete high-throughput subcloning services.