Preparation of Growth Media for Arabidopsis
Arabidopsis Growth Media
Basic Medium
Add the following to deionized water (~ 20% less than final volume)
Final concentrations 500 mL 1 L
Murashige & Skoog Salts†, 1X 2.15 g 4.3 g
Sucrose, 1% 5.0 10.0
MES, 0.05% 0.25 0.5
Adjust to pH 5.7 with KOH.
Adjust to final volume.
Add to bottles with agar: 4.0 g (split in 2 bottles)
Autoclave on liquid cycle 30-40 minutes.
Remove promptly from autoclave; swirl to dissolve agar.
Cool in 55°C water bath until comfortable to the touch.
Pour plates in sterile tissue culture hood.
Hygromycin selection
Add to cooled Basic Medium just before pouring plates (in sterile tissue culture hood):
Stock solutions (filter sterilized) For 500 mL medium
Hygromycin*, 50 mg/mL 300 µL
Carbennicilin*, 50 mg/mL 500 µL
Swirl to mix.
Kanamycin selection
Add to cooled Basic Medium just before pouring plates (in sterile tissue culture hood):
Stock solution (filter sterilized) For 500 mL medium
Kanamycin*, 50 mg/mL 500 µL
Swirl to mix.
DEX induction
Add to cooled Basic Medium just before pouring plates (in sterile tissue culture hood):
Stock solution (filter sterilized) For 500 mL medium
Dex*, 10 mM 5 µL
Swirl to mix.
Basta selection
Prepare exactly as Basic Medium with the following recipe:
Final concentrations 500 mL 1 L
Murashige & Skoog Salts†, 0.5X 1.08 g 2.15 g
Sucrose, 0.5% 2.5 5.0
MES, 0.05% 0.25 0.5
Continue with Basic Medium procedure….
Add to cooled medium just before pouring plates (in sterile tissue culture hood):
Stock solution (filter sterilized) For 500 mL medium
Glufosinate†, 50 mg/mL 500 µL
Swirl to mix.
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