User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/12/18

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18/12/2012

RFP Plate Reader Serial Dilution

  1. Transfer 100 μl of RFP from stock to 100μl DI water. (50% solution)
  2. Transfer 100 μl of 50% solution to 100 μl DI water. (25% solution)
  3. Transfer 100 μl of 25% solution to 100 μl DI water. (12.5% solution)
  4. 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
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