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DSMZ培养基383组分A基础/DSMZ383组分A基础
Desulfobacterium Medium Base,DSMZ medium 383 Solution A Base,DSMZ Medium 383/DSMZ Medium383/DSMZ 383/DSMZ383
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配套试剂
- 1. A1031 - 丙酮酸钠/焦葡萄酸钠 Sodium pyruvate
- 2. A3006 - 无水碳酸钠 Sodium carbonate anhydrous
- 3. SD029 - 微量元素溶液SL-10/微量元素溶液SL10 Trace elements solution SL-10/Trace elements solution SL10
- 4. SD052 - 亚硒酸盐-钨酸盐溶液 Selenite-tungstate solution
- 5. SD104 - 10%硫化钠溶液（0.5g/支） 10% Na2S solution
- 6. SD092 - Wolin氏维生素溶液(10x) Wolin's vitamin solution(10x)
产品详情
培养基配方（每升）
Na2SO4 3.0g
KH2PO4 0.2g
NH4Cl 0.3g
NaCl 21.0g
MgCl2·6H2O 3.0g
KCl 0.5g
CaCl2·2H2O 0.15g
Sodium resazurin(刃天青钠)0.0005g
KH2PO4 0.2g
NH4Cl 0.3g
NaCl 21.0g
MgCl2·6H2O 3.0g
KCl 0.5g
CaCl2·2H2O 0.15g
Sodium resazurin(刃天青钠)0.0005g
使用说明
称取本品28.15g于956ml去离子水中，加入微量元素溶液SL-10(SD029)和亚硒酸盐-钨酸盐溶液(SD052)各1支，搅拌溶解，通入80%H2和20%CO2的混合气体不低于30分钟使pH低于6.0，装入厌氧容器中，121℃灭菌15分钟，冷却，备用。
制备DSMZ培养基383时，另制备并加入溶液B(1g碳酸钠、20ml水，121℃灭菌15分钟)、溶液C(2.5g丙酮酸钠、20ml水，过滤除菌)、溶液D(1支SD092)以及溶液E(取4ml SD104)，混合，使用无菌的酸碱溶液调节pH至7.0-7.2，备用
制备DSMZ培养基383时，另制备并加入溶液B(1g碳酸钠、20ml水，121℃灭菌15分钟)、溶液C(2.5g丙酮酸钠、20ml水，过滤除菌)、溶液D(1支SD092)以及溶液E(取4ml SD104)，混合，使用无菌的酸碱溶液调节pH至7.0-7.2，备用
特殊说明
383: DESULFOBACTERIUM MEDIUM
1. Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at
least 30 min), then distributed in anoxic cultivation vials and autoclaved under the same
gas atmosphere. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas
atmosphere. Solutions C and D are prepared under 100% N2 gas and filter-sterilized.
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. Final pH of the medium should be 7.0 – 7.2.
2. Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution
freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the
beginning. For transfers use 5 - 10% inoculum. Incubate all strains in the dark.
For DSM 2056: Replace pyruvate with 0.70 g/l Na-butyrate, 0.30 g/l Na-caproate, and 0.15
g/l sodium octanoate.
For DSM 3383: Replace pyruvate with indole as substrate. Dissolve 0.30 g of indole in 90
ml water by heating and shaking in a closed bottle under 100% N2 gas atmosphere,
autoclave, then add 7.00 ml of a sterile anoxic stock solution of NaCl (30% w/v) and 0.70
ml of a sterile anoxic stock solution of MgCl2 x 6 H2O (40% w/v). Store the indole-salt
solution in the dark. Reheat and shake before use. Add to sterile medium 30.00 ml/l of the
indole-salt solution in the beginning, and 2 x 30.00 ml/l during growth.
For DSM 3384: The vitamin solution of medium 120 is replaced with 1.00 ml/l of seven
vitamins solution of medium 503. Replace pyruvate with 0.40 g/l Na-benzoate and add
10.00 ml/l clarified rumen fluid (see medium 1310) as growth supplement. Initiation of
growth requires addition of freshly prepared sodium dithionite.
For DSM 4661: Replace pyurvate with resorcinol. During growth the culture is fed once
with the same amount of resorcinol.
For DSM 5091, DSM 9788: Replace pyruvate with 2.00 g/l malonic acid.
For DSM 7044, DSM 7120, DSM 7467, DSM 12567, DSM 13228, DSM 23484: Supplement
medium with 1.00 ml/l seven vitamins solution (see medium 503). Replace pyruvate with
0.40 g/l benzoate and 0.10 g/l yeast extract sterilized separately by filtration and add to
the autoclaved medium from anoxic stock solutions.
For DSM 8540: Replace pyruvate with 0.30 g/l p-hydroxybenzoate.
For DSM 9705: Replace pyruvate with 1.00 g/l Na-glycolate sterilized by filtration.
For DSM 12861, DSM 12883: Replace pyruvate with 1.00 g/l valeric acid.
For DSM 12888: Replace pyruvate with 1.10 g/l Na-butyrate and 1.10 g/l caproic acid.
For DSM 13036: Replace pyruvate with 1.00 g/l betaine.
For DSM 14454: Naphthalene dissolved in heptamethylnonane can be used as substrate
instead of pyruvate.
For DSM 15576, DSM 16219: Replace pyruvate with 1.00 g/l Na-caprylate and supplement
medium with 1.00 ml/l seven vitamins solution (see medium 503) added to the autoclaved
medium from an anoxic stock solution sterilized by filtration.
For DSM 17291: Replace pyruvate with 3.00 g/l Casamino acids.
For DSM 17477: Replace pyruvate with 1.90 g/l sodium glutamate monohydrate and 1.00
g/l yeast extract.
For DSM 21856: Replace pyruvate with 0.90 ml/l 1-butanol.
For DSM 28890: Replace pyruvate with 0.50 g/l yeast extract and 0.68 g/l sodium formate.
For DSM 100305, DSM 105015: Change amount of pyruvate to 2.20 g/l and add 0.10 g/l
yeast extract.
Solution A
Solution B
Solution C
Solution D
Wolin's vitamin solution (10x) 1.00 ml
Solution E
Selenite-tungstate solution (from medium 385)
Trace element solution SL-10 (from medium 320)
First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally
make up to 1000.00 ml.
Wolin's vitamin solution (10x) (from medium 120)
|
Solution A Solution B Solution C Solution D Solution E |
952.00 20.00 20.00 1.00 10.00 |
ml ml ml ml ml |
least 30 min), then distributed in anoxic cultivation vials and autoclaved under the same
gas atmosphere. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas
atmosphere. Solutions C and D are prepared under 100% N2 gas and filter-sterilized.
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. Final pH of the medium should be 7.0 – 7.2.
2. Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution
freshly prepared under N2 and filter-sterilized) may stimulate growth of some strains at the
beginning. For transfers use 5 - 10% inoculum. Incubate all strains in the dark.
For DSM 2056: Replace pyruvate with 0.70 g/l Na-butyrate, 0.30 g/l Na-caproate, and 0.15
g/l sodium octanoate.
For DSM 3383: Replace pyruvate with indole as substrate. Dissolve 0.30 g of indole in 90
ml water by heating and shaking in a closed bottle under 100% N2 gas atmosphere,
autoclave, then add 7.00 ml of a sterile anoxic stock solution of NaCl (30% w/v) and 0.70
ml of a sterile anoxic stock solution of MgCl2 x 6 H2O (40% w/v). Store the indole-salt
solution in the dark. Reheat and shake before use. Add to sterile medium 30.00 ml/l of the
indole-salt solution in the beginning, and 2 x 30.00 ml/l during growth.
For DSM 3384: The vitamin solution of medium 120 is replaced with 1.00 ml/l of seven
vitamins solution of medium 503. Replace pyruvate with 0.40 g/l Na-benzoate and add
10.00 ml/l clarified rumen fluid (see medium 1310) as growth supplement. Initiation of
growth requires addition of freshly prepared sodium dithionite.
For DSM 4661: Replace pyurvate with resorcinol. During growth the culture is fed once
with the same amount of resorcinol.
For DSM 5091, DSM 9788: Replace pyruvate with 2.00 g/l malonic acid.
For DSM 7044, DSM 7120, DSM 7467, DSM 12567, DSM 13228, DSM 23484: Supplement
medium with 1.00 ml/l seven vitamins solution (see medium 503). Replace pyruvate with
0.40 g/l benzoate and 0.10 g/l yeast extract sterilized separately by filtration and add to
the autoclaved medium from anoxic stock solutions.
For DSM 8540: Replace pyruvate with 0.30 g/l p-hydroxybenzoate.
For DSM 9705: Replace pyruvate with 1.00 g/l Na-glycolate sterilized by filtration.
For DSM 12861, DSM 12883: Replace pyruvate with 1.00 g/l valeric acid.
For DSM 12888: Replace pyruvate with 1.10 g/l Na-butyrate and 1.10 g/l caproic acid.
For DSM 13036: Replace pyruvate with 1.00 g/l betaine.
For DSM 14454: Naphthalene dissolved in heptamethylnonane can be used as substrate
instead of pyruvate.
For DSM 15576, DSM 16219: Replace pyruvate with 1.00 g/l Na-caprylate and supplement
medium with 1.00 ml/l seven vitamins solution (see medium 503) added to the autoclaved
medium from an anoxic stock solution sterilized by filtration.
For DSM 17291: Replace pyruvate with 3.00 g/l Casamino acids.
For DSM 17477: Replace pyruvate with 1.90 g/l sodium glutamate monohydrate and 1.00
g/l yeast extract.
For DSM 21856: Replace pyruvate with 0.90 ml/l 1-butanol.
For DSM 28890: Replace pyruvate with 0.50 g/l yeast extract and 0.68 g/l sodium formate.
For DSM 100305, DSM 105015: Change amount of pyruvate to 2.20 g/l and add 0.10 g/l
yeast extract.
Solution A
|
Na2SO4 KH2PO4 NH4Cl NaCl MgCl2 x 6 H2O KCl CaCl2 x 2 H2O Trace element solution SL-10 Selenite-tungstate solution Sodium resazurin (0.1% w/v) Distilled water |
3.00 0.20 0.30 21.00 3.00 0.50 0.15 1.00 1.00 0.50 950.00 |
g g g g g g g ml ml ml ml |
|
Na2CO3 Distilled water |
1.00 20.00 |
g ml |
|
Na-pyruvate Distilled water |
2.50 20.00 |
g ml |
Wolin's vitamin solution (10x) 1.00 ml
Solution E
|
Na2S x 9 H2O Distilled water |
0.40 10.00 |
g ml |
|
NaOH Na2SeO3 x 5 H2O Na2WO4 x 2 H2O Distilled water |
0.50 3.00 4.00 1000.00 |
g mg mg ml |
|
HCl (25%) FeCl2 x 4 H2O ZnCl2 MnCl2 x 4 H2O H3BO3 CoCl2 x 6 H2O CuCl2 x 2 H2O NiCl2 x 6 H2O Na2MoO4 x 2 H2O Distilled water |
10.00 1.50 70.00 100.00 6.00 190.00 2.00 24.00 36.00 990.00 |
ml g mg mg mg mg mg mg mg ml |
make up to 1000.00 ml.
Wolin's vitamin solution (10x) (from medium 120)
|
Biotin Folic acid Pyridoxine hydrochloride Thiamine HCl Riboflavin Nicotinic acid Calcium D-(+)-pantothenate Vitamin B12 p-Aminobenzoic acid (DL)-alpha-Lipoic acid Distilled water |
20.00 20.00 100.00 50.00 50.00 50.00 50.00 1.00 50.00 50.00 1000.00 |
mg mg mg mg mg mg mg mg mg mg ml |
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