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497. CLOSTRIDIUM KN MEDIUM (HYDROXYBENZOATE)

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 Solution A 922 ml - -
2 Solution B 30 ml - -
3 Solution C 1 ml - -
4 Solution D 10 ml - -
5 Solution E 10 ml - -
6 Solution F 10 ml - -
7 , might need strain adjustment 3 g/l - -
Instructions:
  1. Solution A is sparged with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and F are autoclaved under 100% N2 gas atmosphere. Solutions D and E are prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 7.1 - 7.4.
  2. Note: Some cultures are shipped in semi-solid medium which stimulates growth at the beginning. For agar stabs 3.00 g/l agar are added to the complete medium from a sterile anoxic stock solution (2% w/v). Upon receipt add anoxically 1 - 2 ml of the recommended freshly prepared liquid medium to the agar tube and incubate for 3 - 5 days. After incubation transfer 0.5 ml of the resulting cell suspension in the liquid phase to tubes with liquid medium.
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.50 g 1.627 11.454
2 0.20 g 0.217 1.594
3 0.25 g 0.271 5.069
4 1.00 g 1.085 18.558
5 0.40 g 0.434 2.134
6 0.50 g 0.542 7.274
7 0.15 g 0.163 1.107
8 Selenite-tungstate solution 1.00 ml - -
9
(0.1% w/v)
0.50 ml 5.4e-4 0.002
10 920.00 ml 996.042 55288.7
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.5 g 50 471.752
2 30.0 ml 998.207 55408.8
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 Trace element solution SL-10 1 ml - -
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 Vitamin solution 10 ml - -
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 DSM 5671 10 ml - -
2 DSM 5672 10 ml - -
3 DSM 5673 10 ml - -
Instructions:
  1. Use substrates depending on organism (see below).
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.4 g 40 166.54
2 10.0 ml 998.207 55408.8

DSM 5671 #

10
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.4 g 40 -
2 10.0 ml 998.207 55408.8

DSM 5672 #

10
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.15 g 15 -
2 10.00 ml 998.207 55408.8
Instructions:
  1. The concentration of 2-hydroxybenzoate in the medium should not exceed 1.5 mmol per liter; feed substrate several times in order to reach a reasonable cell density.

DSM 5673 #

10
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.75 g 75 543.002
2 10.00 ml 998.207 55408.8
Solution occurs in 161 other media (first defined in medium 385 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 0.5 g 0.5 12.501
2 3.0 mg 0.003 0.011
3 4.0 mg 0.004 0.012
4 1000.0 ml 998.207 55408.8
Solution occurs in 366 other media (first defined in medium 141 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 2.0 mg 0.002 0.008
2 2.0 mg 0.002 0.005
3 10.0 mg 0.01 0.049
4 5.0 mg 0.005 0.015
5 5.0 mg 0.005 0.013
6 5.0 mg 0.005 0.041
7 5.0 mg 0.005 0.01
8 0.1 mg 1.0e-4 6.3e-5
9 5.0 mg 0.005 0.036
10 5.0 mg 0.005 0.024
11 1000.0 ml 998.207 55408.8
Solution occurs in 239 other media (first defined in medium 320 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
2
(25%; 7.7 M)
10.0 ml 3.17 86.943
3 1.5 g 1.5 7.545
4 70.0 mg 0.07 0.514
5 100.0 mg 0.1 0.505
6 6.0 mg 0.006 0.097
7 190.0 mg 0.19 0.799
8 2.0 mg 0.002 0.012
9 24.0 mg 0.024 0.101
10 36.0 mg 0.036 0.149
11 990.0 ml 988.225 54854.8
Instructions:
  1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 ml.

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 7.3
Equipment needed: Autoclave, Filter, Hungate tubes

Associated strains

All strains for this medium: 3
Strains with modifications: 3

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Distilled water 980.23 54411
Na2CO3 1.5 14.153
Na2SO4 1.5 10.56
NaCl 1 17.111
KCl 0.5 6.707
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Cultivation metadata from 3 strains BacDive

phyla 2 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 3 strains

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