Second Selection (WMS-H30) Medium
| Reagent | Weight/Volume |
|---|---|
| MS basic salts | 4.6 g |
| Modified MS vitamin stock (1,000×) | 1 mL |
| L-Glutamine | 500 mg |
| Casein hydrolysate | 100 mg |
| MgCl2·6H2O (250 mg/mL) | 3 mL |
| Maltose | 40 g |
| MES | 1.95 g |
| ddH2O | |
| Adjust pH to 5.8 | |
| Agarose (type I) | 5 g |
| Make up the volume to 1 L, and autoclave at 121°C for 20 min | |
| Cool to 50°C, and then add the reagents below | |
| 2,4-D (1 mg/mL) | 0.5 mL |
| Picloram (2.2 mg/mL) | 1 mL |
| Ascorbic acid (100 mg/mL) | 1 mL |
| Carbenicillin (250 mg/mL) | 1 mL |
| AgNO3 (100 mM) | 50 μL |
| Hygromycin (50 mg/mL) | 0.6 mL |
Pour 25 mL of medium into a 90-mm petri dish, and air-dry in the laminar flow clean bench for 1 h. Seal with parafilm and store at 20–25°C in the dark. The medium can be stored for up to 3 weeks.