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What can I expect to see in a well on a 96 well plate after transfecting the NS0 host cells? |
| | Cells will grow in the wells for up to 2 weeks (utilising available glutamine) and eventually form a lawn covering approximately 80% of the surface of the well. This lawn of cells will then begin to die, the dying cells appearing shrunken and more granular than the healthy NS0 cells. By 3-4 weeks it may be possible to see the first transfectants growing through. Colonies should be visible with the naked eye when the plates are examined from underneath. The colony should appear as a ball of cells and the cells within it should appear quite large, bright and round, and quite distinct from the background of dead cells. This ball of cells should be dispersed (with a pasteur pipette) before it gets too large, as viability will be affected if they remain in the ball. To judge when to disperse the colony, look for signs of the medium starting to change to orange/yellow – the colony should be easily visible with the naked eye before dispersing. |
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