In what areas is pcr testing applied

Molecular biology is considered cutting-edge technology in the current field of medical research, and 44PCR detection technology is one of them, which is widely used in clinical medicine. 44PCR detection technology is mainly used to analyze gene sequences and detect the occurrence of tumors, cancer cells and genetic diseases. Its characteristics are that it can detect mutated genes in a short period of time, can also trace complex genetic organizations, and is easy to combine with other genetic testing methods, so it is widely used in the field of medical research.

Application of PCR in mutation detection of genes:

1. Direct PCR detection of point mutations

1. Direct detection of deletions using PCR:

When there is a deletion within a gene, a pair of primers can be designed on both sides of the deleted fragment using the known DNA sequence of the gene. PCR is then performed and the product is subjected to agarose gel electrophoresis and stained with ethidium bromide. The presence of specific amplification products is detected under an ultraviolet detector. This makes it very easy to determine whether there is a missing DNA fragment in the sample to be tested.

1. Detection of deletion using a pair of primers PCR. If the DNA sequence of a gene is clear and the deletion site is relatively fixed, a pair of suitable primers can be synthesized on both sides of the deletion fragment according to the DNA sequence of the deletion region for PCR. Then, agarose gel electrophoresis and ethidium bromide staining are performed, and the presence of specific amplified fragments is observed under short-wave ultraviolet light. This method is the fastest way to detect gene fragment deletions.

2. Multiplex PCR detection with multiple primer pairs is missing.

3. Detection of loss of heterozygosity using PCR technology.

2. Direct detection of single alkali displacement PCR

1. Direct detection of changes in restriction endonuclease cleavage sites - PCR product enzymatic analysis.

2. 3'-specific PCR, amplification blocking mutation system and allele-specific PCR.

3. 3.PCR direct sequencing.

4. PCR-oligonucleotide probe dot hybridization (PCR-ASb).

2. Rapid screening of mutant genes using PCR technology

The methods mentioned above can directly detect the mutation site, and the prerequisite is that the nature and location of the mutation must be clear. However, in many cases, the site of the gene mutation is not fixed, and the nature is not very clear. Therefore, some fast and simple screening methods are needed to determine whether there is a gene mutation in the sample. In recent years, the rapid mutation screening technology based on PCR technology has been widely used in the detection of gene mutations in tumors and genetic diseases.

1. PCR-single-strand conformation polymorphism

(ii) Detection of gene mutations using denaturing gradient gel electrophoresis (DGGE), constant temperature gel electrophoresis (CGGE) and temperature (TGGE).

In summary, there are many gene mutation detection technologies based on PCR technology, and each has its own advantages and disadvantages, but the most widely used ones are PCR-SSCP, PCR-ASO and PCR cycle direct sequencing. With the continuous emergence of new methods, the research on gene mutations will be greatly promoted.