20.109(F20):M1D2
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Part 3: Seed cells for gamma-H2AX assay
- Obtain one 12-well plates from the front laboratory bench.
- Clearly label the plate with the date and your team information. Include the name of the cell line seeded!
- Carefully place a coverslip into each of the wells EXCEPT C1 and C2 (the bottom row, column 1 and 2).
- Add 2 mL of fresh media to each well that contains a coverslip.
- Calculate the volume of cells that contains 25,000 cells for each cell suspension (25,000 cells / well).
- Use the density you calculated above to determine the volume.
- Mix the cells and add the appropriate volume of cells into each of the wells.
- CHO cells settle quickly in conical tubes. It's important you mix before adding cells to the 12 well plate.
- To ensure that the cells are evenly dispersed in the well, slide the plate up / down and left / right 3-5 times.
- Carefully move your plates to the 37°C incubator
- Clean out the tissue culture hood:
- Aspirate any remaining cell suspensions.
- Dispose of all vessels that held cells in the biohazard waste box and be sure that all sharps are in the sharps jar.
- Remove any equipment or supplies that you transferred into the hood and return to the appropriate location.
- Please leave the equipment that was already there.
- Spray the TC hood surface with 70% ethanol and wipe with paper towels.
- Be sure the paper towels are disposed of in the biohazard waste box!
- Empty the benchtop biohazard bucket into the biohazard waste box.