PCR

PCR laboratory:

     In principle, the amplification testing laboratory is divided into four separate working areas, reagent storage and preparation area, specimen preparation area, and amplification product analysis area. To avoid cross-contamination, access to each work area must strictly follow a single direction.

     Reagent preparation area→sample configuration area→amplification area→product analysis area. The transfer of reagents and samples between each experimental area should be carried out through the transfer window.

Reagent preparation area:

     The main operations in this experimental area are the preparation of storage reagents, the sub-packaging of reagents and the preparation of the main reaction mixture. Reagents and materials used for sample preparation should be shipped directly to this area and must not pass through other areas. The reagent raw materials must be stored in this area, and the required storage reagents must be prepared in this area. There are no strict requirements for the control of airflow pressure in this area.

Sample preparation area:

The main operation in this area is the preservation of samples. Nucleic acid (RNA, DNA) extraction, storage and addition to amplification reaction tubes and determination of DNA synthesis.

 The pressure gradient in this area is required to be positive pressure relative to the adjacent area to avoid aerosol pollution entering this area from the adjacent area. In addition, since contamination by aerosols may occur during sample addition operations, unnecessary movement in this area should be avoided.

Amplified region:

The main operation performed in this region is DNA amplification. In addition, the addition of the prepared DNA template (from the sample preparation area) and the preparation of the reaction mixture from the main reaction mixture (from the reagent storage and preparation area) can also be performed in this area.

The pressure gradient in this area is required to be negative pressure relative to the adjacent area to avoid aerosol leakage from this area. To avoid pollution caused by aerosols, unnecessary movement in the area should be minimized. Individual operations such as sample addition should be carried out in a clean bench.

Product analysis area:

    The main operation in this region is the determination of amplified fragments.

   This area is the main source of contamination of amplification products. Therefore, the pressure gradient in this area is required to be negative pressure relative to the adjacent area, so as to avoid the diffusion of amplification products from this area to other areas.