PCR cloning technology can amplify trace amounts of DNA, generating millions of copies of a specific DNA sequence. PCR is highly sensitive, extremely specific, and high-yield, while also being easily reproducible, fast, and convenient, making it an amazingly powerful tool in molecular biology. With the rapid development of modern life science, PCR cloning technology has been more and more widely applied to various fields in biological research, medical research, virus detection, and the food industry.
PCR cloning Technology Applied to Gene Cloning
Gene cloning and Subcloning via PCR technology plays a significant role in cell biology research. PCR technology can generate millions of copies of a single-copy gene, amplifying a specific DNA fragment that might only be a few picograms. Compared to other gene cloning techniques, PCR omits several tedious processes for cloning of a particular gene fragment from genome DNA, such as enzyme digestion, connection, transformation, DNA library construction, gene screening, gene identification, and Subcloning.
PCR Cloning Technology Applied to DNA Recombination
In molecular biology, PCR cloning technology can be used to construct recombinant DNA molecules by inserting different sources of specific genes or DNA fragments into viruses, plasmids or other vectors in vitro. Recombinant DNA molecules are then imported into reporter cells to amplify and reproduce. After screening, the daughter cells that contain the target gene are further multiply to extract a large amount of DNA. Recombinant DNA technology can be applied to the Human Gene Project, valuable protein expression, gene diagnosis and therapy, genetic modification of animals and plants, and other research fields.
PCR Cloning Technology for Gene Quantification
PCR cloning technology can be applied to quantitatively determine the copy number of a target gene in a sample. The target gene and a single copy reference gene are placed in a tube for PCR cloning. The PCR product is then separated by electrophoretic separation technology and band intensity is observed. Alternatively, the 5’ end of the primer can be marked by a radionucleotide, after which the gene copy number can be determined by radioactivity quantification. PCR cloning technology can also be applied to quantitatively analyze mRNA and tRNA. It can even detect 1TIRNA, which is hard to detect even by Northern blot.
Alteration of endogenous genes and invasion of foreign genes can be threatening to human health. Regardless of whether or not pathogenesis is caused by genetic changes, as long as there is a pathogen, its corresponding existing nucleic acid can be found. With the development of PCR cloning technology and related technologies, PCR can be applied to infectious disease pathogenesis detection and diagnosis, tumor related gene detection, hereditary disease early diagnosis, bone marrow transplant HLA – D locus matches, and evolutionary theory analysis.
Synbio Technologies’s Syno® 2.0 platform can clone a target gene to any specific point on provided vectors without depending on restriction enzyme sites, and can quickly and cost-effectively fulfill a wide variety of client requests for myriad applications in synthetic biology.