After the PCR amplification is completed, do not bring the tube containing the amplified product into the PCR preparation area
●Designate the area where the PCR system is prepared to minimize the entry and exit of items.
●Wear gloves when working in this area and change them frequently. Wear a mask and headgear to reduce the pollution of facial skin and hair cell sources.
●Establish a set of reagents and disposable supplies (including PCR tubes, mineral oil, wax beads, etc.) for individual use. Use new glassware, plastic utensils, and pipettes that have not been exposed to the DNA environment of the laboratory. Use these items to prepare and store solutions.
●Before opening the centrifuge tube containing PCR reagents, use a small centrifuge in the laminar flow cabinet to centrifuge briefly (10s) to centrifuge the liquid to the bottom of the tube. This can reduce the contamination of gloves and pipettes.
●Dilute the DNA template on your own laboratory bench, and bring only the diluent needed to be used into the PCR preparation area.
●After completing the PCR amplification, do not bring the tube containing the amplified product into the PCR preparation area, but perform the subsequent steps on your own laboratory bench.
●PCR-grade water is necessary for all PCR reactions, does not contain ions, salts, nucleases, and has a neutral pH. It can be purchased from Ambion and other suppliers. Removal of contamination of solutions and equipment. The reagents (buffer and water without dNTPs) are irradiated with 254nm wavelength ultraviolet light to inactivate the contaminated DNA.
Ultraviolet radiation can cause adjacent pyrimidine bases in double-stranded DNA to form dimers, so that contaminated DNA can no longer become a template for PCR reactions. Using a commercial UV crosslinker (such as Stratalinker, Agilent products), the solution in the transparent white centrifuge tube can easily be irradiated (200~300mJ/cm2 for 5~20min).
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