培養基配方
Final pH: 7.2 - 7.4
Final volume: 1003 ml
Solution A 942.00 ml
Solution B 30.00 ml
Solution C 20.00 ml
Solution D 1.00 ml
Solution E 10.00 ml
Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C and D are prepared under 100% N2 gas and sterilized by filtration. Solution E is autoclaved under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.2 - 7.4, if necessary.
For DSM 5847: Replace D-glucose with 1.50 g/l of Na (D/L) 3 hydroxybutyrate or 2.00 g/l D-fructose as substrate.
For DSM 5849: Replace D-glucose with 2.50 g/l of Na2 succinate as substrate.
For DSM 5885: Replace D-glucose with 1.60 g/l Na2-maleate as substrate added from an anoxic stock solution sterilzed by filtration. Adjust pH of complete medium to 6.7 - 6.8.
For DSM 6779: Replace D-glucose with 2.00 g/l of xylose or xylan as substrate.
For DSM 10092: Omit glucose. Add 0.70 g/l Na2SO4 to solution A and use 1.50 g/l Na
-propionate as substrate. Reduce the amount of sulfide to 0.10 g/l and use 25 - 50 mg
sodium dithionite per liter (e.g. from a freshly prepared filter-sterilized 5% w/v solution) for
reduction of the medium prior to inoculation.
For DSM 11045: Omit solution C. Use as final concentration: 10 mM taurine, 60 mM Na
-formate, and 200 μM 1,4-naphthochinone as substrates added from anoxic stock solutions sterilized by filtration
For DSM 11046: Add 1.70 g/l NaNO3 to solution A and replace D-glucose with 1.25 g/l taurine as substrate.
For DSM 11261: Replace D-glucose with 1.00 g/l yeast extract and 2.00 g/l Na glycolate as substrates.
For DSM 11262: Replace D-glucose with 1.00 g/l yeast extract and 2.50 g/l Na (D/L)-3 hydroxybutyrate as substrates.
For DSM 11263, DSM 11489: Replace D-glucose with 0.82 g/l Na acetate and 3.20 g/l Na2 fumarate as substrates added from anoxic stock solutions sterilized by filtration.
For DSM 11270: Replace D-glucose with 2.50 g/l taurine as substrate.
For DSM 11480, DSM 12595: Reduce amount of D-glucose to 1.80 g/l.
For DSM 12018: Replace D-glucose with 0.20 g/l yeast extract and 2.66 g/l L-aspartic acid as substrates added from anoxic stock solutions sterilized by filtration.
For DSM 13305: Replace D-glucose with 1.00 g/l yeast extract and 5.00 g/l D-fructose as substrates added from anoxic stock solutions sterilized by filtration.
For DSM 14424, DSM 28450, DSM 100378, DSM 103557, DSM 105239: Supplement medium with 2.00 g/l yeast extract added from an anoxic stock solution sterilized by filtration.
For DSM 15206: Replace D-glucose with 2.00 g/l yeast extract and 2.00 g/l Trypticase peptone (BD BBL) as substrates.
For DSM 15970: Supplement medium after autoclaving with 0.6 g/l syringic acid added from a sterile anoxic stock solution neutralized with NaOH. Replace solution D with 1 ml/l Wolin's vitamin solution (10x). Omit D-Glucose.
For DSM 15978, DSM 27305: Supplement medium with 20 ml/l FeSO4 x 7 H2O solution
(0.1% w/v), 1 ml/l Na-dithionite solution (5% w/v), 0.8 ml/l Methanol, and 2 g/l
Trimethylamine-HCl. Omit D-Glucose.
For DSM 19598: Reduce amount of glucose to 2 mM (0.36 g/l); defined coculture with Methanospirillum hungatei. Incubate in the dark. Anaerobic
For DSM 24856: Omit glucose. Add 0.85 g/l NaNO3 to solution A. Use 0.10 g/l yeast extract and 1.10 g/l Na-pyruvate as substrates. Na-pyruvate is added to the autoclaved medium from a sterile anoxic stock solution sterilized by filtration.
For DSM 25964: Replace D-glucose with 0.50 g/l Na-pyruvate as substrate added from an anoxic stock solution sterilized by filtration. Supplement solution A with 2.0 g/l yeast extract.
For DSM 27520: Omit sodium carbonate, reduce amount of D-glucose to 2.00 g/l and add 1.00 g/l yeast extract as substrate. Adjust pH of complete medium to 5.5.
For DSM 27582: Supplement medium after autoclaving with 0.1 g/l yeast extract, 2.0 g/l D-mannose, and 1 ml/l Wolin's vitamin solution (10x) added from anoxic stock solutions
sterilized by filtration. Omit D-Glucose.
For DSM 100618: Reduce glucose to 3g/l. Add 1g/l yeast extract.
For DSM 101182: Supplement medium after autoclaving with 0.50 g/l yeast extract and 0.40 g/l L-cysteine HCl x H2O added from sterile anoxic stock solutions.
For DSM 103526: Reduce amount of D-glucose to 0.40 g/l and add 1.00 g/l yeast extract as substrate.
For DSM 111565: Replace D-glucose with 0.50 g/l yeast extract and 1.00 g/l L-rhamnose as substrates added from anoxic stock solutions sterilized by filtration .
For DSM 112739: Omit sodium resazurin. Replace D-glucose in solution C with 2.20 g/l sodium phosphite hydrate, and sulfide in solution E with 0.3 g/l DTT (DL-Dithiothreitol).
For DSM 121889, DSM 121891, DSM 121892: Supplement medium with 0.5 g/l cysteineHCl x 2 H2O. Omit Na2S x 9 H2O.
Solution A
KH2PO4 0.20 g
NH4Cl 0.25 g
NaCl 1.00 g
MgCl2 x 6 H2O 0.40 g
KCl 0.50 g
CaCl2 x 2 H2O 0.15 g
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Sodium resazurin (0.1% w/v) 0.50 ml
Distilled water 940.00 ml
Solution B
Na2CO3 1.50 g
Distilled water 30.00 ml
Solution C
D-Glucose 5.00 g
Distilled water 20.00 ml
Solution D
Seven vitamins solution 1.00 ml
Solution E
Na2S x 9 H2O 0.30 g
Distilled water 10.00 ml
Trace element solution SL-10 (from medium 320)
HCl (25%) 10.00 ml
FeCl2 x 4 H2O 1.50 g
ZnCl2 70.00 mg
MnCl2 x 4 H2O 100.00 mg
H3BO3 6.00 mg
CoCl2 x 6 H2O 190.00 mg
CuCl2 x 2 H2O 2.00 mg
NiCl2 x 6 H2O 24.00 mg
Na2MoO4 x 2 H2O 36.00 mg
Distilled water 990.00 ml
First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
Selenite-tungstate solution (from medium 385)
NaOH 0.50 g
Na2SeO3 x 5 H2O 3.00 mg
Na2WO4 x 2 H2O 4.00 mg
Distilled water 1000.00 ml
Seven vitamins solution (from medium 503)
Vitamin B12 100.00 mg
p-Aminobenzoic acid 80.00 mg
D-(+)-biotin 20.00 mg
Nicotinic acid 200.00 mg
Calcium pantothenate 100.00 mg
Pyridoxine hydrochloride 300.00 mg
Thiamine-HCl x 2 H2O 200.00 mg
Distilled water 1000.00 ml
