TISSUE CULTURE STOCK SOLUTIONS AND MEDIA

TISSUE CULTURE STOCK SOLUTIONS AND MEDIA

MS MEDIUM FOR ARABIDOPSIS

To 990 ml H2
O 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)
MgSO47H2
O.......... 15.0  ml  100 mM (24.65 g/L)
CaCl2
7H2
O.......... 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
FeSO47H2
O .........  1.0  ml         (2.780 g/100 ml)
Na2EDTA ...........  1.0  ml  100 mM (3.720 g/100 ml)
MnSO44H2
O..........  1.0  ml  100 mM (1.690 g/100 ml)
ZnSO44H2
O..........  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)
CuSO45H2
O .........  0.01 ml   10 mM (0.170 g/100 ml)
Na2MoO42H2
O .......  0.1  ml   10 mM (0.242 g/100 ml)
CoCl2
6H2
O .........  0.01 ml   10 mM (0.238 g/100 ml)

B5 MEDIUM FOR ARABIDOPSIS

To 990 ml H2
O 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)
NaH2PO4H2
O......... 11.0  ml  100 mM
MgSO47H2
O.......... 10.0  ml  100 mM (24.65 g/L)
(NH4)2SO3.......... 10.0  ml  100 mM
CaCl2
7H2
O..........  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)

MnSO44H2
O..........  0.6  ml  100 mM (1.690 g/100 ml)
ZnSO44H2
O.......... 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)
CuSO45H2
O .........  2.3  ml   10 mM (0.170 g/100 ml)
Na2MoO42H2
O .......  1.0  ml   10 mM (0.242 g/100 ml)
CoCl2
6H2
O .........  1.0  ml   10 mM (0.238 g/100 ml)