1 Mix components thoroughly, adjust pH to 6.0 with 1 N HCl and autoclave under a H2-CO2 (4:1, v/v) gas mixture. After cooling, aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized): | |||||
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NaCl | 15.00 | g | ||
|
MgCl2 x 6 H2O | 4.00 | g | ||
|
KCl | 0.33 | g | ||
|
CaCl2 x 2 H2O | 0.33 | g | ||
|
(NH4)2SO4 | 0.50 | g | ||
|
FeCl2 solution | 1.00 | ml | ||
|
Trace element solution | 1.00 | ml | ||
|
Selenite-tungstate solution | 0.50 | ml | ||
|
Distilled water | 860.00 | ml | ||
|
KH2PO4
(3.3%) |
10.00 | ml | ||
|
Trace vitamins
(*) |
10.00 | ml | ||
|
NaHCO3
(*, 8%) |
10.00 | ml | ||
|
(w/v) MES (pH 6.0, 20%) |
10.00 | ml | ||
|
Iron(III) citrate solution | 100.00 | ml | ||
2 Readjust pH to 6.0 to 6.3, if necessary. |
Compound | Amount | Unit | |
---|---|---|---|
|
HCl
(7.7 M, 25%) |
10.0 | ml |
|
FeCl2 x 4 H2O | 1.5 | g |
|
Distilled water | 990.0 | ml |
Compound | Amount | Unit | |
---|---|---|---|
|
ZnCl2 | 70 | mg |
|
MnCl2 x 4 H2O | 100 | mg |
|
H3BO3 | 6 | mg |
|
CoCl2 x 6 H2O | 190 | mg |
|
CuCl2 x 2 H2O | 2 | mg |
|
NiCl2 x 6 H2O | 24 | mg |
|
Na2MoO4 x 2 H2O | 36 | mg |
|
Distilled water | 1000 | ml |
Compound | Amount | Unit | |
---|---|---|---|
|
NaOH | 0.4 | g |
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
Distilled water | 1000.0 | ml |
Compound | Amount | Unit | |
---|---|---|---|
|
Biotin | 2.0 | mg |
|
Folic acid | 2.0 | mg |
|
Pyridoxine hydrochloride | 10.0 | mg |
|
Thiamine HCl | 5.0 | mg |
|
Riboflavin | 5.0 | mg |
|
Nicotinic acid | 5.0 | mg |
|
Calcium pantothenate | 5.0 | mg |
|
Vitamin B12 | 0.1 | mg |
|
p-Aminobenzoic acid | 5.0 | mg |
|
Lipoic acid | 5.0 | mg |
|
Distilled water | 1000.0 | ml |
1 Mix 2.4 g iron(III) citrate and 90 ml distilled water, and boil until dissolved. Cool to an ambient temperature and adjust pH to 6.0 with 10 N NaOH solution. Fill up to 100 ml with distilled water. |