Day of the experiment, sterilize with bleach again via the above protocol
Sterilize septa at sampling port
Using a pipette, place a single drop of bleach on the septa through the sampling port and allow to sterilize during vial bleaching before experiment
Sterile DI water (or Milli Q water) in an eVOLVER bottle
4 inch needle
50mL syringe
Number of seconds pumped in pump commands are for a 30mL reservoir and 10mL lagoon. If you change these volumes, make sure to define your own number of seconds pumped.
As always, any spills on the min-eVOLVER should be immediately cleaned
If you think something got on an electronic part, unplug the min-eVOLVER, wipe, and allow to dry
Fill both reservoir and lagoon with bleach over efflux line
Remove vials from Smart Sleeves to keep an eye on fluid levels
python3 send_command.py 5555 pump 0,0,20,15,0,0
Check fluid levels
Ensure all tubing is filled with bleach
Sterilize at least 30 minutes
Do not allow bleach to sit in vials for more than a day
Use the below protocol to replace bleach with sterile DI water
Remove bleach from vials
Insert a 4 inch needle and 50mL syringe through the sampling port
Try and get almost all of the bleach
Avoid scratching your vial with the needle as this can change the OD calibration
Hook up sterile DI water bottle to influx
Fill both reservoir and lagoon with sterile DI water over efflux line
python3 send_command.py 5555 pump 0,0,70,60,0,0
Remove vials from Smart Sleeves to keep an eye on fluid levels
Vials can now be safely stored for more than one day without bleach corrosion
Avoid adding too much bleach as it will run down the sides of the vial and change the OD calibration
Flush vial with sterile media using the Remove Bleach protocol
Follow experiment setup
Starting an Experiment
You should have completed both setup and calibrations before proceeding to an experiment.
Highly recommended:
Read the main pages for helpful information not listed here and an example experimental workflow.
Run an experiment with water before doing so with cells, especially if you change the code
Questions?
Ask about experiments in the relevant category on the .
Setup
You will have one command line window for each server and each experiment running. It can also be helpful to have a separate window for send_commands.py
For example for two min-eVOLVER experiments:
2 server windows (server environment)
Use send_command.py to fill the vials to appropriate volumes
Do not inoculate cells until after starting the experiment if you want accurate readings. There is innate variability in the OD of the vials (among other things) and we will blank to your vials during experiment start.
Use send_command.py to set to correct temperature
A calibrated temperature command is in your calibration file temperature_calibration.xlsx
Running the Experiment
Make sure that your eVOLVER computer is set to never sleep. If your computer falls asleep while you're running an experiment, you will not be collecting data during that time. See more info .
In a separate window from your server command line, follow the command line start .
If running more than one min-eVOLVER at a time: you need a separate experiment folder and command line window for each min-eVOLVER (and different port number)
An example fluidics layout for basic chemostat or turbidostat. If necessary, inducer can be programmed to be controlled via the low flow-rate pumps (not shown).
Vials set up for a basic chemostat or turbidostat.
