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- Cut microbore tubing into 48'' lengths, one per port. If a dial-a-wave device is used, 96" lines are necessary to compensate for the increased height of the linear actuators
- Remove the plunger from a 10ml syringe
- Note: For dial-a-wave devices, 25ml or 60ml syringes are used for the media reservoirs. The protocol is the same, but the larger syringes are required to reduce the impact of media depletion.
- Attach a luer stub to the syringe body
- Insert the luer stub into one end of the microbore tubing
- Insert a straight pin into the other end of the microbore tub
- Decant 10ml of sterile water into the syringe, letting it run down the side of the syringe body
- Flick the luer stub to clear any bubbles
- Let the water flow through the tube until all of the bubbles have been cleared
- Top the syringe with a square of parafilm
- Use the pin to poke a hole in the parafilm, and store the tube until it is time to load the chip
- Repeat steps 2 to 10 for remaining water syringes and media syringe(s)
- Once the cells have reached OD600 of ~0.1, transfer 10ml of cells to a 15ml conical tube. Spin at ~1500xg (3500RPM in a Sorvall SS-34 rotor) for 5 minutes.
- While the cells are spinning down, turn on the fluorescence source for the microscope so that the bulb can reach operating temperature.
- Note: During the first 5-10 minutes of on-time, the intensity of the fluorescence source will fluctuate until the bulb reaches operating temperature. Therefore, it is important to allow a sufficiently long warm up time. Once the source is on, leave the output at 0\% 0% to prevent accidental exposure when configuring the run.
- Decant and discard the supernatant. Add 10ml fresh LB media with antibiotics and resuspend the cells by vortexing briefly
- Repeat steps 2 to 10 with the cell syringe
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