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Molecular biology laboratory (PCR) design requirements

Molecular Biology Laboratory (PCR) Design Requirements

PCR Laboratory of molecular biology, also known as gene amplification laboratory. PCR is referred to as the polymerase chain reaction (polymerase chain reaction), is a molecular biology technique, for amplification of specific DNA fragments, and they can be regarded as a special DNA replication in vitro. Through DNA tracking system, can quickly grasp the content of virus in patients with, its accuracy up nanometer level.

PCR laboratory has no strict purification requirements, but in order to avoid the possibility of cross contamination between the various experimental areas, it is appropriate to use the full flow of air distribution. At the same time, strictly control the delivery, exhaust air ratio in order to ensure the pressure requirements of the experimentation area.

1. Reagent Storage and Preparation Area: The experimental area of operation for the storage of reagent preparation, preparation of reagents packaging and the main reaction mixture. Reagents and for specimen of the material should be transported directly to the District, not by other regions. Reagent raw materials must be stored in this area, and in the preparation into the required storage reagent. Control of air pressure, the district did not strict requirements.

2. Specimen Preparation Area: The region of operation for preservation of clinical specimens, nucleic acid (RNA, DNA) extraction, storage and added to the amplification reaction tube and determination of RNA cDNA synthesis. The pressure gradient requirements: relative to the adjacent area for positive pressure, to avoid entering the aerosol pollution in this area from adjacent areas. In addition, since may caused by aerosol pollution occurs during the loading operation, it should be to avoid unnecessary walk in this area.

3. Amplification Reaction Mixture Preparation and Amplification Region: Caused by the leakage of reagent and preparation for operation in this region of DNA or cDNA amplification. In addition, preparation of DNA template and synthesis of cDNA (from the sample area) to join the main reaction mixture (from storage and preparation area) preparation of reaction mixture, also can be in the area. In the nested PCR assay, usually in the first round of amplification must turn on the tube, so a nested PCR had a higher risk of contamination, the second sample must be in the area. The area of pressure gradient requirements: with respect to the adjacent area of negative pressure in order to avoid aerosols from the area. In order to avoid aerosol pollution, should try to reduce in Unnecessary walk in the area. The individual operation, such as adding samples should be carried out in the ultra clean table.

4. Amplified Product Analysis Area: Operations in the region mainly for the determination of amplified fragment. Such as the use of full automatic close to detection and analysis instrument, this area may not be set up. This area is the main amplification product of the sources of pollution, so requirements of the pressure gradient: relative to the adjacent area is negative, to avoid amplification products from the region to spread to other areas.