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Introduction
You will be provided with the HEK293/COS cells that you transfected on Monday. Working in groups of two, take the two samples transfected by your group and assay for luciferase activity. Note, in all cases, the total amount of DNA used for the transfection is 2.0µg.
Luciferase assay
1 x CCLR (cell lysis reagent) Luciferase Assay Reagent
Experimental procedure
2. Add a minimal volume of 1 x CCLR to cover the cells. (250 µl for a 60 mm dish) and incubate for a short period at room temperature.
4. Mix 20 µl of room temperature cell extract with 100 µl room temperature Luciferase Assay Reagent. Place the reaction in a luminometer.
6. Record data
Results Ð transfected cells
Discussion
Controls
