Callus-inducing medium

6-Benzylaminopurine

2,4-Dichlorophenoxyacetic acid (2,4-D)

6-(γ,γ-Dimethylallylamino)-purine (2iP)

Indole-3-acetic acid (IAA)

6% KH₂PO₄

1N KOH

1X MS salts (GIBCO)

Phytagel (Sigma)

Sucrose

500X Vitamix

For 1 liter of medium, dissolve 1X MS salts and 30 g of sucrose in water. Adjust the pH to 5.9 with 1 N KOH. Add 3 ml of 6% KH₂PO₄ and 2.2 g of Phytagel (Sigma).

Phytagel rather than agar is critical for success.

Autoclave the solution. Add 2 ml of 500X Vitamix, 3 mg of IAA, 0.15 mg of 2,4-D, 0.6 mg of 6-benzylaminopurine and 0.3 mg of 2iP.