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Growing Arabidopsis Roots
Arabidopsis roots grown in hydroponic conditions
The reference is Gibeaut, et al. (1997), Plant Physiol. 155, 317-319.
The published protocol (above) was followed strictly except that the bubbling is continuous and that we wet the top of the rockwoll with water, each day until the plants are big enough to have roots crossing the cylinder of rockwool. Also medium was renewed each 15 days.
Arabidopsis roots grown in MS medium
Arabidopsis roots can grown in liquid medium (dependently from external carbon source). I usually use 250-ml flask containing 100-ml of growing medium which is constituted of MS + 30g/l sucrose + 1 µM of indolebutyric acid. Flask containing seeds (around 10 per flask) are growing in continuous darkness (wrapped in aluminum foil), on a rotary shaker (100 rpm), and at room temperature. After 3 weeks of culture, the medium were renewed for an additional week to sustain an exponential growth and finally obtain about 15-g of FW.
Arabidopsis seeds were sterilized as indicated on the following URL:
http://www.its.caltech.edu/~plantlab/protocols/ sterprot.html (which is copy-paste below):
Supplies: 70% EtOH, 100% Bleach, Sterile Water, sterile Eppendorf tubes.
1. Put seeds in an Eppendorf tube.
2. Add 200 uL of 70% EtOH to tube.
3. Immediately add 600 uL of water.
Do only 2 tubes at once since the alcohol treatment is harsh and might affect germination if it is left on the seeds too long.
4. Immediately suck off 600 uL of the supernatant.
5. Repeat steps 3 and 4 four times.
6. Set aside tubes and repeat steps 1 to 5 for all the remaining tubes.
7. Add 200 uL of bleach to each tube.
8. Let the tubes sit for 5 minutes.
9. Add 600 uL of water
10. Suck off 600 uL of the supernatant.
11. Repeat steps 9 and 10 four to five times.