——Enter the laboratory, wear experimental clothes, and wear goggles and other safety protection when necessary.

——Take good care of the experimental equipment and standardize the operation. Students need to operate and use various instruments and equipment under the guidance of their tutors.

——Do not touch, play with or taste experimental reagents (Forbidden) at will, so as to ensure safety and avoid cross contamination of reagents.

——After the experimental operation, clean up the table, and put the reagents and consumables in place.

——Do not shout loudly, and prohibit chasing and fighting in the laboratory, so as to create a quiet and safe experimental environment.

——It is forbidden to eat or drink water in the experimental area, and colored drinks are also forbidden to be brought into the experimental area.

——Due to the impact of the epidemic, masks must be worn when entering the laboratory.

——The strain we used in this experiment is the Escherichia coli strain (DH5α）。

——Plasmid vectors used: GV248, virus packaging auxiliary plasmids (helper1.0, helper2.0).

——At the same time, we also used some experimental reagents: 1KB DNA Marker, normal 250bp-ii DNA ladder, restriction endonuclease of NEB, Taq plus DNA polymerase of Vazyme, T4 DNA ligase, high sugar DMEM medium, fetal bovine serum (FBS), ECL luminous solution color development kit, puromycin, penicillin-streptomycin, reverse transcription kit, RT-PCR kit, CCK-8, temozolomide.
These experimental materials are allowed to be used in level 1 laboratories after testing.
Among them, lentivirus GV248 was used to establish cell lines. GV248 is derived from HIV. It is the second-generation product of the lentivirus vector, which includes three independent vectors: packaging plasmid (pspax2), envelope plasmid (pmd2.G), and transfer plasmid (lenticas9-mCherry). For the sake of experimental safety, we directly choose to ask the company for lentivirus packaging.