MediaDive DSMZ
Content
Media Solutions Ingredients Strains Steps Gas
Search
Medium finder Solution finder Taxonomy search Isolation sources
Tools
Medium builder Unit converter Compare media Compare solutions Prediction SOON
DSMZ

DSMZ 1630e . Synthetic Crenarchaeota Medium (SCM)

What is this page?
Equipment needed: Autoclave Filter
Final pH: 7.6
Select a strain (optional):

Basic salt water medium #

ml
Compound Amount Unit
NaCl 26.0 g
MgSO4 x 7 H2O 5.0 g
MgCl2 x 6 H2O 5.0 g
CaCl2 x 2 H2O 1.5 g
KBr 0.1 g
NaHCO3 buffer (1M) 2.0 ml
HEPES buffer solution 10.0 ml
KH2PO4 solution (2.93 nM) 5.0 ml
Trace element solution (Nitrososphaera) 1.0 ml
FeNaEDTA solution (7.5 mM) 1.0 ml
NH4Cl (1 M) 2.0 ml
Double distilled water 1000.0 ml
1 Incubate all used glass ware and magnetic stirring rods in 0.1 M HCl and rinse 3x with double distilled water to remove residual detergents (always check visual for soap bubbles).
2 Autoclave the basic salt water medium and cool to room temperature. This medium should be free of any precipitations!
3 Aseptically, add per L of basic salt water medium the following solutions from sterile stocks (do not store complemented SCM): Trace element solution (Nitrososphaera), FeNaEDTA solution, NaHCO3 solution, HEPES buffer solution, NH4Cl solution, KH2PO4 solution
4 Adjust pH to 7.6.

HEPES buffer solution #

1000
Compound Amount Unit
NaOH
(pellets) 		24.0 g
HEPES
(free acid) 		238.4 g
Distilled water 1000.0 ml
1 Fist dissolve NaOH in ca 600 ml water, then add HEPES, adjust pH to 7.5 and fill up to 1000 ml volume.

Trace element solution (Nitrososphaera) #

1000
Compound Amount Unit
HCl
(12.5M) 		8 mL
H3BO3 30 mg
MnCl2 x 4 H2O 100 mg
CoCl2 x 6 H2O 190 mg
NiCl2 x 6 H2O 24 mg
CuCl2 x 2 H2O 2 mg
ZnSO4 x 7 H2O 144 mg
Na2MoO4 x 2 H2O 36 mg
Double distilled water 1000 ml
1 This solution contains no iron! Sterilise by autoclaving or filtration (0.2 µm) and store in the dark at 4 °C.

FeNaEDTA solution (7.5 mM) #

1000
Compound Amount Unit
FeNa-EDTA 2753 mg
Double distilled water 1000 ml
1 Sterilise by filtration (0.2 µm) and store in the dark at 4 °C.

NaHCO3 buffer (1M) #

50
Compound Amount Unit
NaHCO3 4.2 g
Double distilled water 50.0 ml
1 Dissolve 4.2 g of NaHCO3 in 50 mL Double distilled water (use a small beaker with stirring rod to dissolve) and sterilize by filtration (0.2 µm) in the laminar flow hood. Store in a sterile 50 mL falcon tube at 4 °C. Avoid high headspace to liquid ratios to ensure stable concentration.

NH4Cl (1 M) #

50
Compound Amount Unit
NH4Cl 2.67 g
Double distilled water 50.00 ml
1 Dissolve 2.67 g ammonium chloride in 50 mL Double distilled water.
2 Filtrate (0.2 µm).

KH2PO4 solution (2.93 nM) #

1000
Compound Amount Unit
KH2PO4 0.4 g
Double distilled water 1000.0 ml

alpha-ketoglutamate solution (20mM) #

1000
Compound Amount Unit
alpha-ketoglutamate 20 mM
Double distilled water 1000 ml