Biological origin mud and water samples
Isolator R.Whittenbury University of Warwick
Isolation Date 01/01/1974
History <- 1974,
Risk group human 1
Recommended growth medium
Medium name Temperature Oxygen relationship Ingredients
Medium 391 37 °C CH4:Air:CO2 45:45:10
dNMS Salt Stock:
MgSO4 x 7H2O 10,0 g
KNO3 10,0 g
CaCl2 x 2H2O 1,5 g
Distilled water 1 L
FeNaEDTA Stock
FeNaEDTA 0,5 g
Distilled water 100 ml
Na2HPO4 Stock
Na2HPO4 x 12H2O 71,7 g
Distilled water 1 L
KH2PO4 Stock
KH2PO4 27,2 g
Distilled water 1 L
1000x trace solution:
Na2EDTA x 2H2O 0,5 g
FeSO4 x 7H2O 0,2 g
H3BO3 0,03 g
CoCl2 x 6H2O 0,02 g
ZnSO4 x 7H2O 0,01 g
MnCl2 x 4H2O 0,003 g
Na2MoO4 x 2H2O 0,003 g
NiCl2 x 6H2O 0,002 g
CuSO4 x 5H2O 2,5 g
Distilled water 1 L
Dilute 20ml of DNMS salts stock to 800ml.
Add 1 ml FeNaEDTA Stock and 1 ml trace solution.
Dissolve and bring to 1 liter (distilled water) For solid media add 15g/l Bacto Agar.
After sterilization, cool to 50°C-60°C and add first 5ml KH2PO4 Stock (filter sterilized) followed by 5ml Na2HPO4 Stock (filter sterilized).
pH of the medium should be 6,8
Medium 399 37 °C CH4:Air:CO2 45:45:10
10x NMS salt stock:
KNO3 10,0 g
MgSO4 x 6H2O 10,0 g
CaCl2 x 2H2O 2 g
Distilled water 1 L
FeEDTA stock
FeEDTA 3,8 g
Distilled water 100 ml
Sodium molybdate stock
Na2MoO4 x 2H2O 0,26 g
Distilled water 1 L
Trace solution:
CuSO4 x 5H2O 1,0 g
FeSO4 x 7H2O 2,5 g
ZnSO4 x 7H2O 2,0 g
H3BO3 0,075 g
CoCl2 x 6H2O 0,250 g
Na2EDTA 1,250 g
MnCl2 x 4H2O 0,1 g
NiCl2 x 6H2O 0,05 g
Distilled water 5 L
Phosphate buffer
Na2HPO4 x 12H2O 71,6 g
KH2PO4 26,0 g
Distilled water 1 L
pH = 6,8
Dilute 100ml of 10x NMS salts stock to 1L.
Add 1 ml of sodium molybdate stock and 1 ml of the trace solution.
Add 0,1 ml of FeEDTA Stock.
Add 15g/l Bacto agar.for plates.
After sterilization, cool enough to hold in the hand and aseptically add 10 ml phosphate buffer (autoclaved separately). If this is done too early the phosphate will precipitate out.
Readily available as gDNA No
