Team:Heidelberg/Project/miRNA Kit
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==Methods== | ==Methods== | ||
- | Dual Luciferase assay | + | Dual Luciferase assay<br> |
We measured the knockdown of firefly luciferase using the Promega Dual Luciferase Reporter Assay. Here, Renilla luciferase is used for normalization. The measurements were conducted on the Promega GLOMAX 96 Microplate Luminometer using the Promega standard protocol. | We measured the knockdown of firefly luciferase using the Promega Dual Luciferase Reporter Assay. Here, Renilla luciferase is used for normalization. The measurements were conducted on the Promega GLOMAX 96 Microplate Luminometer using the Promega standard protocol. | ||
Twenty hours after transfection, cells were washed with 1x PBS and lysed using 1x Passive Lysis Buffer (5x stock solution diluted with distilled water), shaking for 30 minutes at 37°C. 10µl of the lysate were transferred to a white microplate (LumaPlate) as required for Luminometer measurements. | Twenty hours after transfection, cells were washed with 1x PBS and lysed using 1x Passive Lysis Buffer (5x stock solution diluted with distilled water), shaking for 30 minutes at 37°C. 10µl of the lysate were transferred to a white microplate (LumaPlate) as required for Luminometer measurements. |
Revision as of 01:21, 24 October 2010
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