Difference between revisions of "Synergy Machine Protocol for Bio113"
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# 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. | # 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. | ||
# If you are measuring GFP and RFP, measure GFP before RFP. | # If you are measuring GFP and RFP, measure GFP before RFP. | ||
| + | # You will want to measure cell density using the spectrophotometer with a wavelength of light set for 600 nm. | ||
# Choose the protocol called '''Bio113 Synthetic Biology Research''' | # Choose the protocol called '''Bio113 Synthetic Biology Research''' | ||
# Place the 96-well plate inside the machine, and be sure to put well A1 near the marker for A1. | # Place the 96-well plate inside the machine, and be sure to put well A1 near the marker for A1. | ||
Revision as of 16:05, 9 August 2022
Synergy Machine Protocol
- All wells should contain exactly 200 µL of liquid. Be sure to include wells containing broth with no cells for negative control.
- 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.
- If you are measuring GFP and RFP, measure GFP before RFP.
- You will want to measure cell density using the spectrophotometer with a wavelength of light set for 600 nm.
- Choose the protocol called Bio113 Synthetic Biology Research
- 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.
