Methods.docx - Materials PBS-Eppendorf...

Materials:
PBS
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Eppendorf tubes
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Hemocytometer
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Inverted microscope
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Marker pen
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24 well plate
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Sterile
Micropipette and tips
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Pipet aid
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Sterile Pasteur pipets
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Falcons
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Complete medium with
serum: e.g., supplemented media with 10% fetal bovine serum (FBS).
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Trypan blue (0.4%)
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Mammalian cell line
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70% ethanol
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Lipofectamine reagentb
Methodology:
First lab
1- Firstly the old medium was discarded from the cells using micropipettes inside the fumehood.
2- 1 ml of PBS was added for washing the cells. 3- The PBS was discarded using micropipette.
4- 1 ml of trypsin was added to the cells. 5- The cells were incubated at 37c and 5% Co2 for 2
min.
6- Centrifugation was performed at 1200 RPM.
7- 1 ml RPMI media was added to the cells.
8- 100 ul of the cells were transferred into eppindorf.
9- 10 ul of trypan blue were added for counting the cells on 10 ul of cells.
10- 40 ul of the cells were transferred to the 24 well plates.
11- 0.5 ml of complete media was added.
12- The well plate was incubated at 37c and 5% Co2.