TISSUE CULTURE STOCK SOLUTIONS AND MEDIA

To 990 ml H₂O add: Sucrose ........... 10.0 g MOPS .............. 0.5 g Agar .............. 8.0 g Adjust pH to 5.7 with 1 M KOH. Autoclave. When cooled add sterile stock solutions of: KNO3............... 20.0 ml 940 mM (95.00 g/L) NH4NO3............. 20.0 ml 1000 mM (80.04 g/L) KH2PO4............. 12.5 ml 100 mM (13.60 g/L) MgSO47H₂O.......... 15.0 ml 100 mM (24.65 g/L) CaCl₂7H₂O.......... 10.0 ml 300 mM (44.00 g/L) Glycine............ 2.67 ml 10 mM Nicotinate......... 0.4 ml 10 mM Thiamin............ 0.02 ml 10 mM PyridoxineHCl...... 0.24 ml 10 mM Myoinositol........ 5.5 ml 100 mM FeSO47H₂O ......... 1.0 ml (2.780 g/100 ml) Na2EDTA ........... 1.0 ml 100 mM (3.720 g/100 ml) MnSO44H₂O.......... 1.0 ml 100 mM (1.690 g/100 ml) ZnSO44H₂O.......... 3.0 ml 10 mM (0.288 g/100 ml) H3BO3 ............. 1.0 ml 100 mM (0.618 g/100 ml) KI................. 0.5 ml 10 mM (0.166 g/100 ml) CuSO45H₂O ......... 0.01 ml 10 mM (0.170 g/100 ml) Na2MoO42H₂O ....... 0.1 ml 10 mM (0.242 g/100 ml) CoCl₂6H₂O ......... 0.01 ml 10 mM (0.238 g/100 ml) B5 MEDIUM FOR ARABIDOPSIS To 990 ml H₂O add: Sucrose ........... 20.0 g MOPS .............. 0.5 g Agar .............. 8.0 g Adjust pH to 5.7 with 1 M KOH. Autoclave. When cooled add sterile stock solutions of: KNO3............... 26.3 ml 940 mM (95.00 g/L) NaH2PO4H₂O......... 11.0 ml 100 mM MgSO47H₂O.......... 10.0 ml 100 mM (24.65 g/L) (NH4)2SO3.......... 10.0 ml 100 mM CaCl₂7H₂O.......... 3.3 ml 300 mM (44.00 g/L) Myoinositol........ 5.5 ml 100 mM Thiamine........... 3.0 ml 10 mM Nicotinate......... 0.8 ml 10 mM PyridoxineHCl...... 0.48 ml 10 mM 10X Micronutrients. 10.0 ml (see below) Sequestrene 330 Fe. 10.0 ml (0.28 g/100 ml) For Callus Inducing Medium (CIM) add: 2,4-D..................... 0.5 ml Kinetin................... 0.05 ml For Shoot Inducing Medium (SIM) add: IAA....................... 0.15 ml 6-dimethylaminopurine..... 5.0 ml 10X Micronutrients (per 100 ml) MnSO44H₂O.......... 0.6 ml 100 mM (1.690 g/100 ml) ZnSO44H₂O.......... 10.07 ml 10 mM (0.288 g/100 ml) H3BO3 ............. 4.9 ml 100 mM (0.618 g/100 ml) KI................. 4.5 ml 10 mM (0.166 g/100 ml) CuSO45H₂O ......... 2.3 ml 10 mM (0.170 g/100 ml) Na2MoO42H₂O ....... 1.0 ml 10 mM (0.242 g/100 ml) CoCl₂6H₂O ......... 1.0 ml 10 mM (0.238 g/100 ml)