Synergy Machine Protocol
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Synergy Machine Protocol
- 1. Open Gen 5 software
- 2. Click on Existing protocol OR move to step 3
- 3. Click on Create New Protocol
- 4. Select Standard Protocol type
- 5. Click on Procedure
- 6. Under Shake, select Orbital Shake for 3 seconds
- 7. Under Read, select Fluorescence from drop down menu
- a. Enter Emission and Excitation wavelengths
- i. If you are reading more than one type of fluorescence, click the appropriate number of types above the wavelength values.
- b. For RFP, select 585 nm (excitation) and 615 nm (emission) for optimal reading (with 100 gain)
- c. For GFP, select ___ (excitation) and ___ (emission) for optimal reading (with 100 gain)
- a. Enter Emission and Excitation wavelengths
- 8. Under Read, select Absorbance from the drop down menu and enter 590 nm
- 9. Once those values have been submitted, drag tasks into proper order. First Shake, then read Fluorescence, then read Absorbance.
- a. If you are measuring GFP and RFP, measure GFP before RFP.
- 10. Click OK, and from the banner menu, select File, New Task, then Read Now (selecting the procedure you created)
- 11. Place the well inside the machine, and Click OK when ready.
- 12. 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.