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Overview of PCR cloning technology

Update time : 2020-08-12

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 Overview of PCR cloning technology
PCR cloning (polymerase chain reaction) technology refers to polymerase chain reaction, also known as in vitro DNA amplification technology.

Traditional methods of DNA amplification:

The traditional method of DNA amplification is molecular cloning, which requires the construction of a vector containing the target gene to be introduced into cells for amplification, and probes for screening, including DNA digestion, ligation, transformation, culture, and probe hybridization techniques. Although there are no technical difficulties, the operation is complicated, the cycle is long, and it is not conducive to popularization.

Advantages of PCR cloning technology:

PCR cloning technology was first proposed by Mullis in the United States in 1983, and polymerase chain reaction, the simple DNA amplification method, was invented in 1985, which meant the true birth of PCR cloning technology. 

As of 2013, PCR cloning has developed to the third generation technology. PCR cloning technology can amplify a small amount of target DNA fragments to millions of times. 

PCR cloning technology is widely used in the fields of microbiology, medicine, agriculture, and archaeology by virtue of its high sensitivity, strong specificity, high yield, good reproducibility, and quickness and simplicity. 

It has been popularized in various laboratories, greatly The traditional molecular cloning technology is simplified, so that researchers can more easily analyze and identify the target gene.

Basic principles of PCR cloning technology:

PCR cloning is based on the use of DNA that denatures and becomes single-stranded at a high temperature of 95°C in vitro. 

At low temperatures (usually around 60°C), the primers and single-strands are combined according to the principle of base complementary pairing, and then the temperature is adjusted to the optimum DNA polymerase. 

At the reaction temperature (about 72°C), the DNA polymerase synthesizes the complementary chain along the direction of phosphoric acid to five carbon sugar (5'-3'). 

The PCR machine based on polymerase is actually a temperature control device, which can well control the denaturation temperature, renaturation temperature, and elongation temperature.

As a basic molecular biology method, PCR cloning technology is widely used in molecular biology applications such as gene cloning, gene recombination, DNA sequence analysis and gene quantitative detection. At the same time, PCR cloning technology is also used in medical tumor-related gene detection, early diagnosis of genetic diseases, etc., providing effective technical means for medical identification and risk analysis.

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

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