Notebook
0508
1. T98G and U118 cells recovery and culture in DMEM
2. DMEM (FBS) cell culture medium preparation
2. DMEM (FBS) cell culture medium preparation
0510-0524
1. transfer the T98G and U118 cells into the fresh DMEM medium containing 50μM and 200μM TMZ, respectively.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
0525-0608
1. Transfer the T98G and U118 cells incubate in TMZ-containing medium into the fresh DMEM medium containing 100μM
and 400μM TMZ, respectively.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
3. screen for TMZ-resistance-related genes in a bioinformatics way
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
3. screen for TMZ-resistance-related genes in a bioinformatics way
0609- 0623
1. transfer the T98G and U118 cells into the fresh DMEM medium containing 200μM and 800μM TMZ, respectively.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
3. designed the siRNA of PDRG1 gene plasmids and send them to the company for plasmids construction and lentivirus package.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
3. designed the siRNA of PDRG1 gene plasmids and send them to the company for plasmids construction and lentivirus package.
0624-0707
1. transfer the T98G and U118 cells into the fresh DMEM medium containing 400μM and 1600μM TMZ, respectively.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
0708-0721
1. transfer the T98G and U118 cells into the fresh DMEM medium containing 800μM and 3200μM TMZ, respectively.
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
3. After the successful preparation of TMZ-resistant cells, the culture is maintained at a low concentration of TMZ (different cell concentrations vary).
4. transfer the cells to a 6-well plate
2. The culture medium containing TMZ is changed every two days, and the same concentration of TMZ is maintained for two weeks.
3. After the successful preparation of TMZ-resistant cells, the culture is maintained at a low concentration of TMZ (different cell concentrations vary).
4. transfer the cells to a 6-well plate
0722-0723
1. transfect the TMZ resistance T98G and U118 cells with lentivirus of PDRG1-siRNA
2. change the culture medium the next day and screened them using purine
3. measure the resistance of TMZ of the transfected cell line
2. change the culture medium the next day and screened them using purine
3. measure the resistance of TMZ of the transfected cell line
0724
1. collect the transfect cells and detect the green fluorescence
2. total RNA extraction of the transfected cells
3. measure the abundance of PDRG1 gene transcripts by qPCR
2. total RNA extraction of the transfected cells
3. measure the abundance of PDRG1 gene transcripts by qPCR
0725
1. measure the expression level of PDRG1 protein by Western blot
