18/12/2012
RFP Plate Reader Serial Dilution
- Transfer 100 μl of RFP from stock to 100μl DI water. (50% solution)
- Transfer 100 μl of 50% solution to 100 μl DI water. (25% solution)
- Transfer 100 μl of 25% solution to 100 μl DI water. (12.5% solution)
- Transfer 100 μl of 12.5% solution to 100 μl DI water. (6.25% solution)
Plate Reader
- Transfer 50 μl of each solution and a control to the Costar 96 clear bottom black side.
- Here is the software setting:
- Fluorescence
- Endpoint
- Full Plate
- Excitation: 540, Emission: 600
- Optics: Top
- Gain: AutoScale
- Light Source: Xenon Flash, Lamp Energy: High
- Read Speed: Normal, Delay: 100 msec, Measurements/Data Point: 10
- Read Height: 9.5 mm
- Click OK and then Run the experiment.
- Export the data to an excel file.
- The software will deduce the control data from the sample data
Result
Dilution |
RFP (AU)
|
100% |
43125
|
50% |
16143
|
25% |
7978
|
12.5% |
4124
|
6.25% |
2718
|
|