Experiment Metadata
Experiment Name:
EIE2
Goal:
This experiment is set-up to test the effect of environmental changes on the metabolome of Arabidopsis grown under standardized conditions
Growth Conditions:
The standardized growth conditions are as follows:
1. Seeds (between 18 and 20) are sown on media in 100 x 100 x 15mm square Falcon Petri Dishes (Becton Dickinson Labware, catalogue #351112). Seeds were arranged on the plates in a single horizontal line at the 1-cm mark from the top of the plate.
2. Each plate contains between 20 and 25-ml of sterile MS media containing 0.1% (w/v) sucrose.
3. Prior to sowing, seeds were sterilized by treating for 1-minute at room temperature with a 300-µl solution of 50% (v/v) ethanol, this solution was removed and replaced with a 300-µl solution consisting of 1% (v/v) Tween 20 (Fischer BioReagents, catalogue #BP33750), and 50% (v/v) bleach solution (Clorox), and incubated at room temperature for 10-minutes. The seeds were then washed with three changes of 0.3-ml of sterile water.
4. Upon sowing with seeds, the plates were wrapped with Micropore tape (3M Health Care, catalogue #1530-0), and then stored horizontally for 4-days at 4 oC, with illumination of 1 µmol/m2.
5. On the 5th day, plates were moved to the growth room, and held in a vertical position in Plexi-glass holders for 16-days - this growth room condition is labeled in Table I as the "normal" growth condition.
6. On the 17th day, at 8am local time, plates were transferred to different growth conditions for a period of 24-hours (treatments in Table I).
7. At the end of this period, Petri plates were opened and the aerial portions of these plants were harvested either immediately upon plate opening or after a 1-h or 3-h delay (Harvest Delay in Table I).
8. Upon harvesting, plant material was quenched by immersion in liquid nitrogen and storage at -70 0C
Timeline
| Event Date | Event |
|---|---|
| 04/25/06 and 04/26/06 | Sowing of Seeds |
| 04/29/06 and 04/30/06 | Move into growth room |
| 05/15/06 and 05/16/06 | Move into treatments |
| 05/16/06 and 3/17/06 | Harvest |
| 05/22/06 | Shipment to analytical labs |