Clostridium pfennigii Medium

[所属分类:培养基配方] [发布时间:2020-5-14] [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

Clostridium pfennigii Medium

培养基配方
Composition per 1001.0mL:
Solution A....................................................................890.0mL
Solution B....................................................................100.0mL
Solution C.....................................................................10.0mL
Solution D......................................................................1.0mL
pH 7.0–7.2 at 25°C
Solution A:
Composition per 890.0mL:
Sodium vanillate.................................................................2.0g
Yeast extract....................................................................2.0g
Resazurin ......................................................................1.0mg
Rumen fluid, clarified........................................................267.0mL
Mineral solution...............................................................50.0mL
Vitamin solution................................................................5.0mL
Trace elements solution SL-10 ..................................................1.0mL
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 890.0mL. Mix thoroughly. Adjust pH to 6.9.
Sparge with 80% N2 + 20% CO2 for 20 min. Distribute 8.9mL into an-
aerobic tubes under 80% N2 + 20% CO2 . Autoclave under 80% N2 +
20% CO2 for 15 min at 15 psi pressure–121°C.
Mineral Solution:
Composition per liter:
KH2PO4 ...........................................................................10.0g
NaCl...............................................................................8.0g
NH4Cl .............................................................................8.0g
MgCl2 ·6H2O.......................................................................6.6g
CaCl2 ·2H2O.......................................................................1.0g
Preparation of Mineral Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl...................................................................6.2mg
Nicotinic acid....................................................................2.5mg
p-Aminobenzoic acid..............................................................1.25mg
Thiamine·HCl ...................................................................1.25mg
Pantothenic acid.................................................................0.62mg
Biotin ..........................................................................0.25mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Adjust pH to 7.0. Mix thor-
oughly.
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 FeCl2 ·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.
Solution B:
Composition per 100.0mL:
NaHCO3 ..........................................................................5.0g
Preparation of Solution B: Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Sparge with 80% N2 + 20% CO2 for 20 min.
Solution C:
Composition per 10.0mL:
L -Cysteine......................................................................0.24g
Preparation of Solution C:  Add  L -cysteine to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Autoclave under
80% N2 + 20% CO2 for 15 min at 15 psi pressure–121°C.
Solution D:
Composition per 1.0mL:
Na2S·9H2O......................................................................78.0mg
Preparation of Solution D:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 1.0mL. Mix thoroughly. Autoclave under
80% N2 + 20% CO2 for 15 min at 15 psi pressure–121°C.
Preparation of Medium: To each tube containing 8.9mL of sterile
solution A, add (using a syringe) 1.0mL of sterile solution B, 0.1mL of
sterile solution C, and 0.01mL of sterile solution D.
Use:  For the cultivation and maintenance of Clostridium pfennigii.