TISSUE CULTURE STOCK SOLUTIONS AND MEDIA MS MEDIUM FOR ARABIDOPSIS

MS MEDIUM FOR ARABIDOPSIS

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)

MgSO₄·7H₂O.......... 15.0  ml  100 mM (24.65 g/L)

CaCl₂·7H₂O.......... 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

FeSO₄·7H₂O .........  1.0  ml         (2.780 g/100 ml)

Na₂EDTA ...........  1.0  ml  100 mM (3.720 g/100 ml)

MnSO₄·4H₂O..........  1.0  ml  100 mM (1.690 g/100 ml)

ZnSO₄·4H₂O..........  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)

CuSO₄·5H₂O .........  0.01 ml   10 mM (0.170 g/100 ml)

Na₂MoO₄·2H₂O .......  0.1  ml   10 mM (0.242 g/100 ml)

CoCl₂·6H₂O .........  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)

NaH₂PO·4H₂O......... 11.0  ml  100 mM

MgSO₄·7H₂O.......... 10.0  ml  100 mM (24.65 g/L)

(NH4)₂SO₃.......... 10.0  ml  100 mM

CaCl₂·7H₂O..........  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

10× 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

10× Micronutrients (per 100 ml)

MnSO₄·4H₂O..........  0.6  ml  100 mM (1.690 g/100 ml)

ZnSO₄·4H₂O.......... 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)

CuSO₄·5H₂O .........  2.3  ml   10 mM (0.170 g/100 ml)

Na₂MoO₄·2H₂O .......  1.0  ml   10 mM (0.242 g/100 ml)

CoCl2·6H2O .........  1.0  ml   10 mM (0.238 g/100 ml)