25th of September protocol

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=25th of September Luciferase assay protocol=
<b>This protocol has been discarded due to interfering of Promega lysis buffer with the protein measurements.</b>

===Cell preparation===
*Pellet 1 ml of grown culture when the culture is around 0.6 by centrifuging 5 min at 10000 rpm in a tabletop centrifuge. 
*Lyse the cells by adding 1x lysis buffer from the Renilla Luciferase Assay kit.
*Freeze the cells at -20 until measurements will be done. 
===Measurements===
*Mix 1 ul of 100X luciferase substrate in 100 ul of assay buffer per sample in the luminometer's reagent tube no. 1
*Put 20 ul of lysed cells in a white 96 wells plate. 
*Put the prime plate in the luminometer (usually on top)
*Put the tube with assay buffer under reagent needle no. 1, make sure the tip of the needle is in a position to reach all the assay buffer. 
*Prime the luminometer with 1000 ul assay buffer in the priming plate(make sure the tubing is rinsed before)
*Measure luciferase activity by: 
**Adding 100 ul of assay buffer to a well
**Wait 2 seconds 
**Integrate luminescence for 10 seconds.
**Repeat for every well
**There is a standard protocol on the computer in which you only have to indicate the wells to be assayed.
*Purge the tubing (Assay buffer can be stored and frozen for short periods (1 week at most) according to the technical manual)
*Rinse the tubing with 5000 ul of ethanol, and purge it.
*Rinse the tubing with 5000 ul of H2O, and purge it. 
*The tubing and injector should be clean and empty now.

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