To begin, navigate to the TIMER2.0 database website interface. Select the Cancer Exploration tab. Input the gene ID into the search box and click Submit to produce the differential expression images of TMEM200A in various tumor types.
For transfection, label three micro-centrifuge tubes as siRNA-1, 2, and 3. Then add transfection reagent followed by the basal medium in each tube. add four micrograms of siRNA to the respective tubes and mix thoroughly.
Now distribute the mixture into the three wells of the six-well plate containing the adenocarcinoma cells. Gently shake the plate to ensure even distribution of the mixture. Label the three wells where the mixture was added as siRNA 1, siRNA two, and siRNA three.
After incubating the cells for four to six hours, replace half of the medium in the labeled wells with fresh complete medium. Incubate the cells for 24 to 48 hours before using them for quantitative RTPCR and Western blotting.
