Reagents preparation area:
● The most “clean” area in labor, existence of nucleic acids is not allowed, including standards and positive controls in the reagents.
● Storage and preparation place for all clean items for experiments such as centrifuge tubes, pipette tips, etc.
● Commercial kit: 75% ethanol, diethyl pyrocarbonate (DEPC) treatment water, organic solvents, etc. may need to be self-prepared
● Self-made reagents: prepare for a larger amount once, divide them in small vials for storage. By every detection, use one small vial , discard the unspent one.
● The instruments and equipment mainly include sample dispensers, balances, centrifuges and refrigerators etc., better to equip with ultra-clean workbench.
Specimen preparation area
● The instruments and equipment mainly include sample mixing equipment, extractor for nucleic acid, bio-security cabinet, sample adding equipment, high-speed desktop (refrigerated) centrifuge, heating modules or water bath, refrigerator etc. Biological safety cabinets should not be placed in the entrance of the laboratory and other places which are easily affected by the movement of people, should not be directly placed opposite to split-type air conditioner.
● Mainly used for the mixing of blood samples and the extraction of nucleic acid samples.
● Pipette tip with a filter element is necessary fine silica sand
● When using a heating module, such as filling fine silica sand in the hole of modules, then incubate the specimen tube in the fine sand, a more consistent incubation temperature can be obtained.
PCR amplification and analysis area
● The preparation of the amplification reaction system and the addition of the extracted nucleic acid can be carried out in the specimen preparation area or in this area. The key is to prevent product contamination. If space permits, it can also be carried out in a separate area.
● When adding samples, add the extracted nucleic acid template samples firstly, close the lid after each addition, then add the positive quality control nucleic acid template, afterwards the sample preparation negative control and only contain the amplification negative control of the main reaction mixture diluted as the sample. The purpose of this is to detect the cross-contamination of the previous amplification products as much as possible.
● Power supply of thermal cycler should be dedicated and equip with an uninterruptible power supply (UPS) or regulated power supply.
● After each amplification, a movable UV lamp can be used to irradiate the amplification thermal cycler.
● Thermal cycler should be calibrated regularly
