BT Medium (DSMZ Medium 816)

[所属分类:培养基配方] [发布时间:2020-3-17] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

BT Medium
(DSMZ Medium 816)

培养基配方
Composition per 1090mL:
NaCl.............................................................1.0g
KCl..............................................................0.5g
MgCl2 ·6H2O.....................................................0.4g
NH4Cl ...........................................................0.3g
KH2PO4 ..........................................................0.2g
CaCl2 ·2H2O....................................................0.15g
Resazurin ......................................................0.5mg
NaHCO3  solution ..............................................50.0mL
Na2S·9H2O solution............................................13.0mL
Hydroxybenzoate solution.......................................10.0mL
Yeast extract solution..........................................5.0mL
Trypticase™ solution............................................5.0mL
Trace elements solution SL-10 ..................................1.0mL
Selenite-tungstate solution.....................................1.0mL
Na2CO3  solution................................................variable
pH 7.6 ± 0.2 at 25°C
Na2S·9H2O Solution :
Composition per 20.0mL:
Na2S·9H2O........................................................0.6g
Preparation of Na2S·9H2O Solution : Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Selenite-Tungstate Solution:
Composition per liter:
NaOH...........................................................0.5g
Na2WO4 ·2H2O..................................................4.0mg
Na2SeO3 ·5H2O.................................................3.0mg
Preparation of Selenite-Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Na2CO3 Solution:
Composition per 100.0mL:
Na2CO3 ............................................................5.0g
Preparation of Na2CO3 Solution:  Add Na2CO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ...........................................................10.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Hydroxybenzoate Solution :
Composition per 10.0mL:
3-Hydroxybenzoic acid...........................................2.8g
Preparation of Hydroxybenzoate Solution : Add 3-hydroxyben-
zoic acid to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Neutralize with NaOH. Filter sterilize.
Trypticase™ Solution :
Composition per 10.0mL:
Trypticase™.....................................................1.0g
Preparation of Trypticase™ Solution : Add Trypticase™ to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Yeast Extract Solution :
Composition per 10.0mL:
Yeast extract...................................................1.0g
Preparation of Yeast Extract Solution:  Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O....................................................1.5g
CoCl2 ·6H2O....................................................190.0mg
MnCl2 ·4H2O ...................................................100.0mg
ZnCl2 ...........................................................70.0mg
Na2MoO4 ·2H2O...................................................36.0mg
NiCl2 ·6H2O.....................................................24.0mg
H3BO3 ...........................................................6.0mg
CuCl2 ·2H2O.....................................................2.0mg
HCl (25% solution)...............................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, Na2S·9H2O solution, yeast extract solution, hydroxybenzoate so-
lution, Trypticase™ solution, selenite-tungstate solution, and Na2CO3
solution, to distilled/deionized water and bring volume to 1.0mL. Mix
thoroughly. Adjust pH to 7.2–7.6. Sparge with 80% N2 + 20% CO2 . Au-
toclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobi-
cally add 50.0mL NaHCO3  solution, 13.0mL Na2S·9H2O solution,
10.0mL hydroxybenzoate solution, 5.0mL yeast extract solution,
5.0mL Trypticase™ solution, and 1.0mL selenite-tungstate solution.
Mix thoroughly. Adjust pH to 7.6 Na2CO3  solution. Aseptically and an-
aerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Sporotomaculum hydroxybenzoicum.