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

Reagent preparation area → sample preparation 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 stored reagents, the packaging of reagents and the preparation of main reaction mixture. Reagents and materials for sample preparation shall be transported directly to this area and shall not pass through other areas. Reagent raw materials must be stored in the area and prepared into the required storage reagents in the area. There are no strict requirements for the control of airflow pressure in this area.

Sample preparation area:

The main operations in this area are the preservation of samples, extraction of nucleic acids (RNA, DNA), storage and addition to amplification reaction tubes, and the determination of DNA synthesis.

The pressure gradient in this area is required to be positive relative to the neighboring area to avoid aerosol pollution entering this area from the neighboring area. In addition, unnecessary movement in the area should be avoided due to the possibility of contamination from aerosols during sampling operations.

Amplification area:

The main operation 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 main reaction mixture (from the reagent storage and preparation area) into the reaction mixture can also be carried out in this area.

The pressure gradient in this area is required to be negative relative to the adjacent area to avoid the leakage of aerosols from this area. To avoid pollution caused by aerosols, unnecessary movement within the area should be minimized. Individual operations such as adding samples should be carried out in the ultra-clean table.

Product analysis area:

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

This area is the most important source of amplification product contamination, so the pressure gradient in this area is required to be negative pressure relative to the neighboring area to avoid the diffusion of amplification products from this area to other areas.