Ingolia Lab Turbidostat

Sterilizing the Media Reservoir

The media reservoir can be sterilized in advance. Once sterilized, the reservoir can also be filled in advance.

1. Place a large stir bar into the media bottle if needed
2. Loosely fit the reservoir cap onto the reservoir bottle and ensure that the media delivery tubing within the bottle reaches the bottom
3. If media delivery manifolds are needed, assemble these onto the media delivery connector.
4. Close the media fill and media delivery connectors with male Luer caps.
5. Close the vent connector with a female Luer cap.
6. Autoclave to sterilize
7. Uncap the vent connector and attach a sterile Luer 13mm disc filter
8. Screw the cap tightly onto the bottle

Sterilizing the Growth Chamber

The growth chamber can be sterilized in advance of an experiment. Once sterilized, the growth chamber can be placed in a warm room in order to equilibrate the temperature prior to use.

1. Fill the humidifier flask with >200 ml deionized water
2. Insert the stopper into the flask and close the air inlet with a female Luer cap
3. Place a stir bar into the growth chamber.
4. Loosely fit the growth chamber cap onto the growth chamber bottle
5. Connect the air outlet from the humidifer to the air (yellow) connector on the growth chamber cap.
6. Close the inoculation (blue) connector on the growth chamber cap with a male Luer cap.
7. Connect the waste line to the waste (red) connector on the growth chamber cap and close the other end of the waste line with a female Luer cap.

8. Connect the shorter end of the media pump line to the media (green) connector on the growth chamber cap. Close the other end of the media line with a female Luer cap.

   

9. Autoclave on liquid cycle to sterilize.
10. Uncap the air inlet connector on the humidifier and attach a sterile Luer 13mm disc filter

11. Screw the growth chamber cap tightly onto the growth chamber bottle

    As the system cools, air should be drawn in through the filter and bubble through the humidifer.