Amplification examination laboratory is generally divided into four separate working areas: reagent storage and preparation area, sample preparation area, amplification reaction mixture preparation and amplification area and amplified product analysis area. In order to avoid cross contamination, all working areas must be entered in a single direction.
Reagent preparation area--sample preparation area--amplification area--product analysis area
Reagent and samples between each experimental area should be passed through the window.
Reagent preparation area
The main operations of the reagent preparation area include preparation/packaging of reagents and main reaction mixture preparation. Reagents and materials used for sample production shall be directly transferred to this area without going through other areas. Reagent raw materials must be stored in this area and prepare the required storage reagents in this area. As for air pressure control, no strict requirements are required.
Sample preparation area
The main operations in the area include sample preservation, nucleic acid (RNA, DNA) extraction, storage and addition to amplification reaction tube and determination of DNA synthesis.
The pressure gradient in this area is required to be positive pressure ralative to the adjacent area. In order to avoid aerosol pollution from the adjacent area. In addition, due to the pollution caused by aerosol in the sample operation, it is necessary to avoid unnecessary movement in this area.
Amplification area
The main operations in this area include DNA amplification. In addition, addition of prepared DNA template (from sample preparation area) and preparation of reaction mixture from main reaction mixture (from reagent storage and preparation area) can also be carried out in this area.
The pressure gradient in this area is required to be negative pressure relative to the adjacent area in order to avoid leakage of aerosol from this area. In order to avoid pollution caused by aerosol, it is necessary to avoid unnecessary movement in this area. Individual operation such as sample addition should be carried out on super clean table.
Product analysis area
The main operations in this area include determination of amplified fragments.
This area is the main source for amplification product contamination, so the pressure gradient in this area is required to be negative pressure relative to the adjacent area in order to avoid amplification product from passing from this area to other areas.
