Daniel:Protocols/SplitCells: Difference between revisions

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>Djacobse
(Created page with "=Splitting Cells= Back to Main ===Protocol=== 1. Check cells for confluency, cells should be at least 70% confluent before splitting cells 2. Aspirate...")
 
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  1. Check cells for confluency, cells should be at least 70% confluent before splitting cells
  1. Check cells for confluency, cells should be at least 70% confluent before splitting cells
  2. Aspirate media, wash with PBS
  2. Aspirate media, wash with 5 mL PBS, aspirate PBS
  3.
3. Add 500 uL Trypsin solution
  4. Incubate 3-5 minutes at 37 C
5. Add 9.5 mL DMEM to dish, pipette several times to resuspend cells
6. Add 2 mL cell suspension to 8 mL DMEM in a new dish, repeat for several dishes if desired
7. Swirl dishes to spread cells, do not spill media
8. Put new dishes back in incubator
9. Rinse old dish with 10% bleach until solution becomes clear and aspirate

Latest revision as of 17:07, 1 October 2013

Splitting Cells[edit]

Back to Main

Protocol[edit]

1. Check cells for confluency, cells should be at least 70% confluent before splitting cells
2. Aspirate media, wash with 5 mL PBS, aspirate PBS
3. Add 500 uL Trypsin solution
4. Incubate 3-5 minutes at 37 C
5. Add 9.5 mL DMEM to dish, pipette several times to resuspend cells
6. Add 2 mL cell suspension to 8 mL DMEM in a new dish, repeat for several dishes if desired
7. Swirl dishes to spread cells, do not spill media
8. Put new dishes back in incubator
9. Rinse old dish with 10% bleach until solution becomes clear and aspirate