Updated January/2023

2 Weeks Prior	1 Week Prior	Experiment Start
- Plant 1 tray of pea plants (one seed per pot) and place in growth chamber in the greenhouse (16:8 light:dark)	- Retrieve 4 plants from the greenhouse growth chambers, place 4 aphids (mature adults without wings) on each plant - Place plants in incubator at 20C (16:8) for 24 hours - After 24 hours, remove adult aphids, leave nymphs to develop for 1 week.	- Collect 2 week old plants from the greenhouse (9-12 plants depending on how many incubators work) - One round tray goes in each incubator - Three plants per tray - One even-aged adult aphid on each plant (from experimental prep)

• 2 weeks prior: Plant 1 tray of pea plants (one seed per pot) and place in growth chamber in the greenhouse

• 1 week prior: 4 plants from the greenhouse growth chambers, 4 aphids (mature adults without wings) on each

• Place plants in incubator 2 at 20C

• Aphids are left on for 24 hours at which point the adults are removed and the plants are returned to the incubator for one week for the next experiment.

• Collect 2 week old plants from the greenhouse (9-12 plants depending on how many incubators work)

• One round tray goes in each incubator

  • Three plants per tray

  • One even-aged adult aphid on each plant (from experimental prep)

• Set up incubators

  • In Celsius:

    • Select "Settings", then "log on device" (device numbers are the same as the incubator number), then "OK" from the menu options

    • File>load tempering profile>from file>select profile>OK

    • Run>start>create log file> name file in the format as others>save

    • Check to make sure it is running, there should be a legend in the top right corner

    • Repeat steps until all incubators are set up

• Remove trays from the incubator and count aphids on each plant

  • Record in lab notebook

  • Update spreadsheet and metafile

• Shut down incubators

  • In Celsius:

    • Run>stop

    • File>load tempering profile>from file>select matching profile>OK

    • File>load protocol data>from file> select matching protocol> OK

    • File>save protocol data> to report>select save to excel file (make sure that all data is in the selected column)...>choose name matching protocol(remove.pro)>OK

  • Excel file should open with data

    • Confirm that temperature data was recorded
    • Add the respective excel protocol files to a new folder and name it
    • Run the cat_stats code to perform a data validation for each temperature regime
    • Update the metafile and aphid spreadsheet with the experimental data, data validation results, and any info on errors (eg. gaps)