When performing PCR operations, the operator should strictly abide by some operating procedures to minimize the possibility of PCR contamination or prevent the occurrence of contamination.1. Divide the operation area: At present, ordinary PCR can not achieve single-person single-tube and complete closed-tube operation. However, whether it can achieve single-person single-tube or not, the experimental operation is required to be carried out in three different areas. Pre-processing and post-processing should be carried out in different isolation areas: (1) Specimen processing area, including preparation of amplification templates. (2) Amplification zone, including preparation of reaction solution and PCR amplification. (3) Product analysis area, gel electrophoresis analysis, product photography and preparation of recombinant clones.

The temperature rise and fall rate, the accuracy and range of temperature control, the sample volume and the hot lid function are undoubtedly the key parameters that users are most concerned about.

Step composition:① Denaturation of template DNA: After the template DNA is heated to about 94℃ for a certain period of time, the template DNA double-stranded or the double-stranded DNA formed by PCR amplification is dissociated to make it single-stranded so that it can be combined with the primer.

Gene amplification is the use of DNA polymerase to carry out specific chain replication, which is a commonly used technique at present. According to the purpose of DNA amplification and testing standards, gene thermal cycler can be divided into four categories: ordinary thermal cycler, gradient thermal cycler, in situ thermal cycler, and real-time thermal cycler.

Placement of the thermal cyclerThe thermal cycler should be placed in a place with low humidity, less dust, far away from water and heat sources, and no corrosive gas or strong magnetic field interference. The ambient temperature should be controlled between 18~35℃.