Methods and materials
Cell culture medium
Making cell culture medium, requires two processes, this includes the medium that has PSG and other without the PSG. 15 ml of 10% FBS was added into a culture flask with 1.5 ml 100x hippos. The culture flask was filled with DMEM up to 150 ml and this will be the medium without the PSG and this process completed the medium without the PSG. The same procedure was followed for the PSG medium, 1.5 ml of PSG was added and that completed the medium with PSG.
Bacterial culture plates for NFkb and GFB
3.5 g of 7 g/250 ml agar and 6.5 g of 25 g/100 ml LB was added to a 500 ml flask. 250 ml of distilled water was added to the same flask. The flask was autoclaved for 20 minutes and it was left to cool down for 10 minutes. The solution was carefully poured into each plate and it was left on the bench for 48 hours in order for the gel to polymerized.
Cell splitting
HeLa cell was vacuumed out of the T-25 flask and 4 ml of PBS was pipetted into the flask to wash cell. PBS was also vacuum out of the R-25 flask and 2 ml of trypsin was pipetted into the T-25 flask. The flask with trypsin was incubated for five minutes for total detachment of the cell that might be inside the flask. One milliliter (ml) of fresh medium with PSG was added to the Trypsin that was …show more content…
500 µL of optiMEM was added into two 1.5 ml micro centrifuge tube. 12 µL of lipofectamin 2000 was added into one tube and 1.5 µL and 1.5 µL of DNA (NFkb) was added into other tube. Both of the micro centrifuge tube was inverted for proper mixture for five minutes. The solution was added into one another after five minutes of inversion, then it was left to stay on the rakes tube for another 20 minutes. The mixture solutions were gently pipetted into the cell that are already inside a six well plate. The six well plates were stored in the incubator for 24 hours at 37°C room
Cell culture medium
Making cell culture medium, requires two processes, this includes the medium that has PSG and other without the PSG. 15 ml of 10% FBS was added into a culture flask with 1.5 ml 100x hippos. The culture flask was filled with DMEM up to 150 ml and this will be the medium without the PSG and this process completed the medium without the PSG. The same procedure was followed for the PSG medium, 1.5 ml of PSG was added and that completed the medium with PSG.
Bacterial culture plates for NFkb and GFB
3.5 g of 7 g/250 ml agar and 6.5 g of 25 g/100 ml LB was added to a 500 ml flask. 250 ml of distilled water was added to the same flask. The flask was autoclaved for 20 minutes and it was left to cool down for 10 minutes. The solution was carefully poured into each plate and it was left on the bench for 48 hours in order for the gel to polymerized.
Cell splitting
HeLa cell was vacuumed out of the T-25 flask and 4 ml of PBS was pipetted into the flask to wash cell. PBS was also vacuum out of the R-25 flask and 2 ml of trypsin was pipetted into the T-25 flask. The flask with trypsin was incubated for five minutes for total detachment of the cell that might be inside the flask. One milliliter (ml) of fresh medium with PSG was added to the Trypsin that was …show more content…
500 µL of optiMEM was added into two 1.5 ml micro centrifuge tube. 12 µL of lipofectamin 2000 was added into one tube and 1.5 µL and 1.5 µL of DNA (NFkb) was added into other tube. Both of the micro centrifuge tube was inverted for proper mixture for five minutes. The solution was added into one another after five minutes of inversion, then it was left to stay on the rakes tube for another 20 minutes. The mixture solutions were gently pipetted into the cell that are already inside a six well plate. The six well plates were stored in the incubator for 24 hours at 37°C room