IGEM:IMPERIAL/2009/Assays Protocols/M3 Assays: Difference between revisions
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ii) Basal levels of GFP expression suggesting that the promoter is not functioning properly. <br> | ii) Basal levels of GFP expression suggesting that the promoter is not functioning properly. <br> | ||
iii) Initial increase in flourescence, followed by a plateauing of the flourescence, as the restriction enzymes destroy the genetic material within the cell. | iii) Initial increase in flourescence, followed by a plateauing of the flourescence, as the restriction enzymes destroy the genetic material within the cell. | ||
==Cell Death Assay== | ==Cell Death Assay== | ||
Revision as of 10:00, 3 August 2009
Assays for Cell Death Strategy
Assay to determine cell death
Restriction Enzyme Assay
Another Possibility
Using GFP marker
Basic genetic circuit for the restriction enzyme testing construct:
Here, there are 3 possibilities for the expression of the GFP, and these are shown below:
i) Non-functional restriction enzyme co-expressed with the GFP.
ii) Basal levels of GFP expression suggesting that the promoter is not functioning properly.
iii) Initial increase in flourescence, followed by a plateauing of the flourescence, as the restriction enzymes destroy the genetic material within the cell.
Cell Death Assay
By Colony Forming Units
1) Plating dilution series on TSB Agar Plates
2) Counting of colonies after overnight incubation
3) Determining the number of colony forming units (number of cells still alive)
