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SetupAttachMachinery

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NEED: Diagrams for attaching the syringe to the pump, diagram for placing culture vessels in their chambers, diagrams for connecting effluent tubes to waste containers

ATTACH SYRINGE TO SYRINGE PUMP

1. Unscrew syringe pump bracing bracket
   • (diagram coming soon)
2. Place syringe connected to 3' tube of octopus tubing system into holder
   • (diagram coming soon)
3. Fasten bracing bracket around the newly inserted syringe
   • (diagram coming soon)
4. Place tubing connected to the syringe between the solenoid pinch valve
   • (diagram coming soon)

PLACE CULTURE VESSELS INTO THEIR CHAMBERS

1. Inside the incubator place each culture vessel into a chamber in order of how they are connected to the octopus tubing system
   • (diagram coming soon)
2. Tape the top of each culture vessel to its chamber
3. Turn on chambers (switch is outside of incubator)

Setup Diagrams
Figure 1: Tubing inserted into pinch valve

Figure 2:Air filters connected to culture vessels

ATTACH MEDIA TUBING TO PINCH VALVES

1. Place the media input tubing that connects to the input ports on the culture vessels into their respective pinch valve
   1. Feed the 2' sections of the octopus lines through the valves around the motor. One at a time, remove the plastic covering piece by sliding it down and off.
   2. Insert the line and slide the plastic piece back on from the bottom.
   3. Culture vessel in chamber 1 should be connected to pinch valve 1 (furthest pinch valve from opening of incubator) and the rest should follow in that order
      • See Figure 1

CONNECT AIR INTAKE TUBING TO AIR PUMPS

1. Connect air intakes to air pumps
   1. Unwrap aluminum foil from a Teflon air filter
   2. Connect previously unwrapped Teflon air filter to tubing hanging from incubator ceiling
      • See Figure 2
   3. Repeat substeps 1-2 for the remaining 7 Teflon air filters
   4. Turn on air pumps (switch is outside of incubator)

CONNECT EFFLUENT TUBING TO WASTE BOTTLES

1. Spray ethanol onto effluent tube holder (each holder takes 4 effluent tubes)
   • (diagram coming soon)
2. Attach effluent tubes to effluent tube holder in order (culture vessel 1 then 2 then 3 etc.)
   1. Unwrap aluminum foil from effluent tube
   2. Spray newly exposed end with ethanol
   3. Insert tube into the effluent tube holder
   4. Repeat substeps steps 1-3 for remaining culture vessels
   • (diagram coming soon)
3. Insert effluent tube holder into a waste bottles
   • (diagram coming soon)

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