Erdschreiber medium

Total volume 1 liter 2 liter

1. Filtered seawater [FSW] 800 ml 1600 ml
2. Distilled water [DW] 200 ml 400 ml
3. NTA [Nitrilotriacetic acid] 0.1 g 0.2 g
4. TRIS Amino methane [CH2N (CH2OH)3] 0.1 g 0.2 g
5. NaNO3 [Sodium nitrate] – 100 mg/ml 1 ml 2 ml
6. Na2HPO412H2O [Disodium hydrogen phosphate] – 120 mg/ml 1 ml 2 ml
7. Vitamin B12, 0.5 g/ml 1 ml 2 ml
8. Thiamine HCl, 200 g/ml 1 ml 2 ml
9. Soil extract 50 ml 100 ml

Before calibration, pH is about 6. Adjust to pH 7.8

Stock solutions:

NaNO3 : 10 g/ 100 ml DW

Na2HPO412H2O : 2 g/ 100 ml DW

Vitamin B12 : 50 mg/ 100 ml, then dilute 1000x to get 0.5 g/ml

Thiamine HCl : 200 mg/ 100 ml

Then preserve the above solutions with 1 drop of n-butylchloride: dichloroethane = 2:1.

Soil extract:

DW: soil = 2:1. Boil the mixture for 2 hours. Leave overnight for the soil to settle down.
Siphon out the extract & centrifuge at 1,000 rpm, 10ºC for 15 minutes. GF/C. Autoclave
at 120ºC for 20 minutes.
SWII medium (1 litre)

1. FSW 1,000 ml
2. TRIS Amino methane [CH2N (CH2OH)3] 500 mg
3. KNO3 [Potassium nitrate] 72 mg
4. KH2PO4 [Potassium dihydrogen phosphate] 4.5 mg
5. C3H7Na2O6P5H2O [Na2- glyserolphosphate disodium salt pentahydrate] 10.5 mg
6. Na2SiO3H2O [Sodium metasilicate] 30 mg
7. Fe-EDTA 3.6 mg
8. Vitamin B12 (from stock solution) 2 g
9. Biotin (from stock solution) 1 g
10. Thiamine HCl (from stock solution) 100 g

Adjust to pH 8.0 – 8.2.

PES medium

i) PES A

1. DW 100 ml
2. NaNO3 [Sodium nitrate] 10 g
3. C3H7Na2O6P5H2O [Na2- glyserolphosphate disodium salt pentahydrate] 1.7 g
4. Thiamine HCl 10 g
5. Vitamin B12 0.1 mg
6. Fe(NH4)2 (SO4)26H2O [Ferrous ammonium sulphate] 0.7 mg
7. H2BO3 [Boric acid] 0.2 g
8. Na2-EDTA 0.3 g

ii) PES B

1. DW 100 ml
2. MnCl2 [Manganese chloride] 4g
3. CaCl2 [Calcium chloride] 0.1 g
4. ZnCl2 [Zinc chloride] 0.4 g
5. FeCl2 [Iron chloride] 1g

iii) PES C

1. DW 100 ml
2. Na2SiO3H2O [Sodium metasilicate] 4.5 g