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Methanothermobacter/Methanobacterium Transduction Agar (DSMZ Medium 863)
[所属分类:培养基配方] [发布时间:2021-5-27] [发布人:网站管理员2] [阅读次数:] [返回]
Methanothermobacter/Methanobacterium
Transduction Agar
(DSMZ Medium 863)
Basal salts solution.................................................................100.0mL
Sodium carbonate solution.......................................................24.0mL
Titanium (III) citrate solution ..................................................16.0mL
Trace elements solution ...........................................................10.0mL
Cysteine solution........................................................................5.0mL
NaS solution...............................................................................5.0mL
pH 7.0 ± 0.2 at 25°C
Sodium Carbonate Solution:
Composition per 100.0mL:
Na 2 CO 3 .......................................................................................10.6g
Preparation of Sodium Carbonate Solution: Add Na 2 CO 3 to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C. Store anaerobically.
Basal Salts Solution:
Composition per liter:
KH 2 PO 4 .......................................................................................68.0g
NH 4 Cl .........................................................................................21.2g
Resazurin .................................................................................10.0mg
Preparation of Basal Salts Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 .
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................9.55g
MgCl 2 ·6H 2 O................................................................................4.06g
FeCl 2 ·4H 2 O.................................................................................0.98g
NaWO 4 ·6H 2 O............................................................................0.264g
NiCl 2 ·6H 2 O...............................................................................0.118g
Na 2 Se 2 O 3 .......................................................................................0.1g
Na 2 MoO 4 ·4H 2 O ........................................................................0.024g
CoCl 2 ·6H 2 O ..............................................................................0.023g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 7.0 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................1.0g
Preparation of Cysteine Solution: Add L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Adjust pH to 10.0. Sparge with 100% N 2 . Store anaerobically.
NaS Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................1.0g
Preparation of NaS Solution: Add Na 2 S·9H 2 O to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to
10.0. Sparge with N 2 . Store anaerobically.
Titanium (III) Citrate Solution:
Sodium citrate, 0.2M................................................................50.0mL
Titanium(III) chloride, 15%.......................................................5.0mL
Preparation of Titanium (III) Citrate Solution: Mix solutions.
Neutralize with sodium carbonate solution. Sparge with N 2 . Store an-
aerobically.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except titanium cit-
rate solution, to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 60°C. Add 16.0mL titanium citrate
solution. Mix thoroughly. Pour into sterile Petri dishes or distribute to
sterile tubes. Store anaerobically.
Use: For the transduction of Methanobacter spp. and Methanobacterium
spp.
Transduction Agar
(DSMZ Medium 863)
山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com
Composition per 1016.0mL:
Agar ............................................................................................15.0gBasal salts solution.................................................................100.0mL
Sodium carbonate solution.......................................................24.0mL
Titanium (III) citrate solution ..................................................16.0mL
Trace elements solution ...........................................................10.0mL
Cysteine solution........................................................................5.0mL
NaS solution...............................................................................5.0mL
pH 7.0 ± 0.2 at 25°C
Sodium Carbonate Solution:
Composition per 100.0mL:
Na 2 CO 3 .......................................................................................10.6g
Preparation of Sodium Carbonate Solution: Add Na 2 CO 3 to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C. Store anaerobically.
Basal Salts Solution:
Composition per liter:
KH 2 PO 4 .......................................................................................68.0g
NH 4 Cl .........................................................................................21.2g
Resazurin .................................................................................10.0mg
Preparation of Basal Salts Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 .
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................9.55g
MgCl 2 ·6H 2 O................................................................................4.06g
FeCl 2 ·4H 2 O.................................................................................0.98g
NaWO 4 ·6H 2 O............................................................................0.264g
NiCl 2 ·6H 2 O...............................................................................0.118g
Na 2 Se 2 O 3 .......................................................................................0.1g
Na 2 MoO 4 ·4H 2 O ........................................................................0.024g
CoCl 2 ·6H 2 O ..............................................................................0.023g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 7.0 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................1.0g
Preparation of Cysteine Solution: Add L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Adjust pH to 10.0. Sparge with 100% N 2 . Store anaerobically.
NaS Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................1.0g
Preparation of NaS Solution: Add Na 2 S·9H 2 O to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to
10.0. Sparge with N 2 . Store anaerobically.
Titanium (III) Citrate Solution:
Sodium citrate, 0.2M................................................................50.0mL
Titanium(III) chloride, 15%.......................................................5.0mL
Preparation of Titanium (III) Citrate Solution: Mix solutions.
Neutralize with sodium carbonate solution. Sparge with N 2 . Store an-
aerobically.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 gas atmosphere. Add components, except titanium cit-
rate solution, to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 60°C. Add 16.0mL titanium citrate
solution. Mix thoroughly. Pour into sterile Petri dishes or distribute to
sterile tubes. Store anaerobically.
Use: For the transduction of Methanobacter spp. and Methanobacterium
spp.
