Synergy Machine Protocol for Bio113

Synergy Machine Protocol

1. All wells should contain exactly 200 µL of liquid. Be sure to include wells containing broth with no cells for your blank.
2. We use the wavelengths of 460 nm (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.
3. If you are measuring GFP and RFP, measure GFP before RFP.
4. You will want to measure cell density using the spectrophotometer with a wavelength of light set for 600 nm.
5. Choose the protocol called Bio113 Synthetic Biology Research
6. Place the 96-well plate inside the machine, and be sure to put well A1 near the marker for A1.
7. 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.