medium no. 12: modified Murashige and Skoog medium, 0.5 mg/L 1-naphthaleneacetic acid, 1 mg/L 6-benzylaminopurine, pH 6.5

Plant cell lines

• rpc00016: Spinacia oleracea Spi-WT cell suspension culture

Chemicals and stock solutions

(All stock solutions are stored at 4°C)

• MS salt mix

  Murashige and Skoog Plant Salt Mixture, FUJIFILM Wako Pure Chemical Corporation (#392-00591)

• Sucrose

• MS_VT

  Chemical	Concentration (mg/mL)
  Nicotinic acid	0.5
  Pyridoxine·HCl	0.5
  Thiamine·HCl	0.1
  Glycine	2

• MS_inositol

  Chemical	Concentration (mg/mL)
  myo-Inositol	40

• NAA (1 mg/mL)

  Chemical	Concentration (mg/mL)
  NAA·K	1.2

  Potassium 1-naphthylacetate, FUJIFILM Wako Pure Chemical Corporation (#161-04021)

• BAP (1 mg/mL)

  Chemical	Concentration (mg/mL)
  6-Benzylaminopurine	1

  Dissolve 6-benzylaminopurine in small volume of KOH (1 N), and fill up with distilled water

• NaOH (1 N)

Medium preparation

1. Dissolve the following chemicals in approximately 800 mL of distilled water.

   Chemical	Amount
   MS salt mix	1 bag (1 L)
   Sucrose	20 g

2. Add following stock solutions, and fill up to approximately 950 mL with distilled water.

   Stock solution	Volume (mL)
   MS_VT	1
   MS_inositol	2.5
   NAA (1 mg/mL)	0.5
   BAP (1 mg/mL)	1

3. Adjust the pH of the solution to 6.5 with NaOH (1 N), and fill up to 1 L with distilled water.

4. Autoclave the medium at 121°C for 20 min.

Formulation

Chemical	Concentration (mg/L)
KNO3	1900
NH4NO3	1650
CaCl2·2H2O	440
MgSO4·7H2O	370
KH2PO4	170
H3BO3	6.2
MnSO4·4H2O	22.3
ZnSO4·7H2O	8.6
KI	0.83
Na2MoO4·2H2O	0.25
CuSO4·5H2O	0.025
CoCl2·6H2O	0.025
FeSO4·7H2O	27.8
Na2-EDTA	37.3
Nicotinic acid	0.5
Pyridoxine·HCl	0.5
Thiamine·HCl	0.1
Glycine	2
myo-Inositol	100
Sucrose	20000
NAA·K	0.6
6-Benzylaminopurine	1