Publish Time: 2022-02-09 Origin: Site
The polymerase chain reaction is a molecular biology technique used to amplify a specific DNA (or RNA) fragment. It can be regarded as a special DNA replication in vitro. The biggest feature of PCR is that it can A substantial increase.
PCR (Polymerase Chain Reaction) uses DNA to become single-stranded when denatured at a high temperature of 95°C in vitro. At low temperature (often around 60°C), the primers and the single-strand are combined according to the principle of base complementary pairing, and then adjusted. The temperature reaches the optimum reaction temperature of DNA polymerase (about 72°C), and DNA polymerase synthesizes complementary strands along the direction of phosphate to five-carbon sugar (5'-3'). The PCR instrument based on polymerase is actually a temperature control device, which can be well controlled between the denaturation temperature, the renaturation temperature and the extension temperature.
A central issue in PCR laboratory design is how to avoid contamination. In practical work, the following types of contamination are common: contamination of amplification products; contamination of natural genomic DNA; contamination of reagents and contamination between specimens. Therefore, to avoid pollution, the first thing to do is to prevent, not to exclude.
The work area is strictly divided, and each experimental area must be clearly marked (such as eye-catching house numbers or different ground colors, etc.) to avoid confusion of equipment, items, reagents, etc. in different experimental areas. Reasonable system settings, reasonable air-conditioning and ventilation system settings, try to use an air-conditioning system with full delivery and full exhaust; strict airflow pressure control to ensure different pressure requirements in different experimental areas.
For standardized operation, the technicians of the clinical gene amplification testing laboratory must undergo induction training. During the experimental operation, the operator must wear gloves and change them frequently. Timely and correct cleaning. The area must be cleaned immediately after the experimental work is over.
Strict management: strictly control the personnel entering and leaving the laboratory. Use work clothes with clearly distinguishable signs (such as different colors) in each experimental area. When the staff leaves, they should not take the work clothes in this area to other areas; try to reduce unnecessary walking in the experimental area to reduce the possibility of cross-contamination sex. The amplification product analysis area is the main source of amplification product contamination. The waste liquid cannot be dumped in the laboratory. It must be immersed in disinfectant and then discarded away from the laboratory. Disposable materials such as used tips should also be used. After being soaked in disinfectant for disinfection, unified treatment, such as incineration, etc.
Laboratory construction is a complex issue. We do not discuss the issue of personnel here. The training and precipitation of laboratory personnel also requires a process. In the process of construction, we must adapt measures to local conditions, do not use rigor, and slowly find out what suits you according to your own reality. Development of construction ideas. Scientific and rational construction of the laboratory, better play the role of the laboratory.