Synergy Machine Protocol for Bio113
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Synergy Machine Protocol
- All Wells should contain 200 µL of liquid. Be sure to include wells containing broth with no cells for negative control.
- We use the wavelengths of 460nm (excitation) and 490 nm(emission) for GFP, and 585 nm(excitation) and 615 nm (emission) for RFP, to minimize noise/interference. However, some of the GFP reading will be from bleeding over of RFP.
- If you are measuring GFP and RFP, measure GFP before RFP.
- Place the 96-well plate inside the machine, and be sure to put well A1 near the marker for A1.
- After the machine has run, you can export the data to Excel using the Excel button on the banner of the window that pops up.
Note- if only a few wells give a value of OVRFLW, try lowering the gain from 100 to 95 or 90 (or lower if necessary). Enter the value on the page with the wavelengths for fluorescence.
