Application of NOB1 gene in treating brain glioma disease
A technique for brain glioma and glioma, applied in the field of research on the pathogenesis of brain glioma, can solve the problem of unclear specific function of NOB1 gene
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[0028] Materials and methods
[0029] cell culture
[0030] HEK293T cells and glioma cells A172 and U373 were purchased from American Type Culture Collection (ATCC). Using DMEM medium, additionally add 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin. Cells were cultured in an incubator at 37°C in a humidified atmosphere with 5% CO2.
[0031] RNA extraction
[0032] Human glioma tissue and normal brain tissue were placed in liquid nitrogen immediately after surgery and stored in a -80°C refrigerator until RNA extraction. Total RNA was extracted according to the operation manual of Trizol reagent. Cell lines A172 and U373 were extracted RNA by the same method.
[0033] RNA reverse transcription and Real-time PCR
[0034] Take 2 μg of RNA to synthesize cDNA according to the operation manual of M-MuLV reverse transcriptase. The synthesized cDNA (20 μg) was mixed with SYBR GreenMasterMix and amplified in the CFX96 real-time monitoring system. Each sam...
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