Be familiar with experimental steps(including the configuration of soft AGAR,western blot) and listed experiment required list
Revived cells,inoculation and expansion
Select an appropriate auxin
July 13th
Configuration reagents used in this experiment
Autoclaving reagent and experiment apparatus
Observing cell metamorphosis
July 14th
Configurate the lower gum of SDS-PAGE
Observing cell metamorphosis
July 15th
Configurate the supernatant gum of SDS-PAGE,extract protein, electrophoresis, trarsmembrane, primary antibody, secondary antibody, visualization and observe
the configuration of soft AGAR
Observing cell metamorphosis
July 16th
Re extracting protein and do western blot
Observing cell metamorphosis
finish the configuration of soft AGAR
Preparation of solid medium, in preparation for the transfection
July 17th
Finished the rest of western blot, got the result of the experience
Collected T293 cells and saving them by freezing
July 18th
Tried to make T protein band by western blot, but it failed
July 19th
Collected protein extraction cancer cells we need urgently
Did western blot twice to test the actin of gastric cancer cell and Hybrid system cancer cells separately
Collected the information of Competent cells
July 20th
Did two western blot experiments, which were 193, 231,hela and hela, 231, to the end of primary antibody incubation
Transformed competent cells, into two boards, PX300F and PX300
July 21th
The transformation of PX300F failed, transformed it again
The transformation of PX300F succeeded, shaked bacteria for the night, prepared extract plasmid
Finished yesterday’s rest of western blot, second incubation resistance enhancement, analyzed the result of the experiment
Observed the cells, collected T293 cells and saved them by freezing
July 22th
Did two western blot. The first experiment, did two pieces of gel to test T protein and KLF4 protein in gastric cancer cell lines. The other one, did two pieces of gel to test T protein and actin in hybrid system cancer cells
Because concentration was too low, the extraction of PX300 plasmid failed. Cultivated cells by shaking cells, prepared to extract plasmids. Coated PX300F plates succeeded, selected cloning cells to cultivate by shaking cells
July 23th
The result of yesterday’s western blot was unsatisfactory, tested partly T protein and actin in hybrid system cancer cells again
Extracted PX300 and PX300F plasmids
July 24th
Finished the rest of yesterday’s western blot, analyzed the result of the experiment