Callus-inducing medium (CIM)

1X Gamborg’s B5 salts

20 g glucose

0.5 g 2-[N-morpholino]ethanesulfonic acid (MES)

KOH (1 N)

7.5 g agar, tissue-culture-grade (for solid medium)

500 μl 2,4-D (2000X)

500 μl kinetin (2000X)

For 1 liter CIM, add Gamborg’s B5 salts, glucose, and MES to H₂O. Adjust pH to 5.7 with KOH. For solid medium, add the tissue-culture-grade agar. After autoclaving, cool medium to ~60ºC and then add the 2,4-D (final concentration 0.5 mg/liter) and kinetin (final concentration 0.05 mg/liter).