Publish Time: 2022-09-06 Origin: Site
The PCR laboratory is divided into four areas. Entry into each working area must be strictly in a single direction. Different working areas use different working clothes (for example, different colors). When staff leave the working areas, they are not allowed to take out their work clothes. The four areas are:
1. Reagent preparation area n
2. Sample processing area n
3. Nucleic acid amplification region n
4. Product analysis area n
If a full-automatic analyzer is used, the regions can be combined appropriately, and the three and four regions can be combined for amplification product analysis.
The functions of each PCR region are:
Reagent preparation area: preparation, sub assembly and storage of amplification reagents.
Sample processing area: sample registration and packaging; Nucleic acid extraction, storage and sampling.
Amplification product analysis area: determination, result analysis, registration and report of amplification products.
The pre amplification area and the post amplification area should be strictly separated, and different rooms should be used. It is better to have a certain interval between the two areas.
A small amount of reagent can be prepared in the sample processing area by using the commercial kit.
On the premise that the following operation and cleaning requirements are met, the sample processing area and reagent preparation area can be set in the same room:
Operate in the biosafety cabinet.
Each experimenter and experimental group used their own reagents and consumables.
Containers containing contaminated materials shall be sealed and disposable.
Use effective methods to clean and disinfect the operation area and shared equipment before and after the experiment.
PCR laboratories can be in the form of dispersion or combination, and now the combination is the mainstream. To complete a set of PCR experiments, reagent preparation, sample processing, nucleic acid amplification and product analysis are usually four experimental processes. If the experimental process requires, the sample crushing process should also be added.
1、 Dispersed form PCR Laboratory
The so-called decentralized PCR laboratory refers to the experimental rooms that complete the above experimental process, which are far away from each other and are distributed. For the PCR laboratory with this arrangement, since each experiment is not easy to interfere with each other, there is no need for special conditions.
2、 Combined form: PCR Laboratory
The so-called combined PCR laboratory refers to the form in which the experimental rooms that complete the four experimental processes of PCR are arranged adjacent to each other to form an independent experimental area. For the combined PCR laboratory, because each laboratory is centrally arranged, it is easy to cause mutual interference. Therefore, there are certain requirements for the overall layout and barrier system. Each room is provided with a buffer room at the entrance to reduce indoor and outdoor air exchange.
The reagent preparation room and sample processing room should be slightly positive pressure to prevent the air containing nucleic acid aerosol from entering and causing pollution. The positive pressure effect can be achieved by controlling the inlet air volume to be larger than the exhaust air volume.
The nucleic acid amplification room and the product analysis room should have a micro negative pressure to prevent the aerosol containing nucleic acid from diffusing out to contaminate the reagent and sample. The negative pressure effect can be achieved by controlling the exhaust air volume to be larger than the inlet air volume.
Under ideal conditions, positive pressure can be set in the buffer room of PCR laboratory, so that indoor air does not flow outdoors and outdoor air does not flow indoors.
The air inlet of PCR laboratory is controlled by the original central air conditioner. It is required to install the air outlet of the central air conditioner at the specified point, and the height is 2600mm after the floor is paved.
If a fluorescent PCR instrument is used, the amplification chamber and the product analysis chamber can be combined.
If the room depth allows, a dedicated corridor inside the PCR can be set. It should be pointed out that the buffer room is more meaningful than the dedicated corridor in reducing indoor and outdoor air exchange.
Each area should have one-way experimental process flow, logistics, people flow and air flow to form a protective barrier of one-way flow, avoid mutual interference between experiments, and prevent nucleic acid aerosol from polluting the experimental process and producing false results.
The walls of the laboratory, including the ceiling, shall be of firm structure and good air tightness; All internal corners should adopt arc-shaped lines for transition; The inner wall of the wall is clean, non adsorption, corrosion-resistant, easy to clean and disinfect; The ground materials shall meet the requirements of no gap, no leakage, smoothness and corrosion resistance.
If a fluorescent PCR instrument is used, the amplification chamber and the product analysis chamber can be combined.
If the room depth allows, a dedicated corridor inside the PCR can be set. It should be pointed out that the buffer room is more meaningful than the dedicated corridor in reducing indoor and outdoor air exchange.
Each area should have one-way experimental process flow, logistics, people flow and air flow to form a protective barrier of one-way flow, avoid mutual interference between experiments, and prevent nucleic acid aerosol from polluting the experimental process and producing false results.
The walls of the laboratory, including the ceiling, shall be of firm structure and good air tightness; All internal corners should adopt arc-shaped lines for transition; The inner wall of the wall is clean, non adsorption, corrosion-resistant, easy to clean and disinfect; The ground materials shall meet the requirements of no gap, no leakage, smoothness and corrosion resistance.