[所屬分類:培養(yǎng)基配方] [發(fā)布時間:2020-7-15] [發(fā)布人:] [閱讀次數(shù):] [返回]

山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Ferulate Medium

培養(yǎng)基配方
Composition per 1016.0mL:
Solution A.............................................................916.0mL
Solution B..............................................................70.0mL
Solution C..............................................................10.0mL
Solution D..............................................................10.0mL
Solution E .............................................................10.0mL
pH 7.2 ± 0.2 at 25°C
Solution A:
Composition per 916.0mL:
Pancreatic digest of casein.............................................1.0g
Resazurin .............................................................1.0mg
Mineral solution.......................................................50.0mL
Rumen fluid, clarified.................................................50.0mL
Vitamin solution........................................................5.0mL
Trace elements solution SL-10...........................................1.0mL
Mineral Solution:
Composition per liter:
Nitrilotriacetic acid...................................................12.5g
NaCl.....................................................................1.0g
FeCl3 ·4H2O.............................................................0.2g
MnCl2 ·4H2O.............................................................0.1g
CaCl2 ·2H2O.............................................................0.1g
ZnCl2 ...................................................................0.1g
CuCl2 ..................................................................0.02g
Na2SeO3 ................................................................0.02g
CoCl2 ·6H2O...........................................................0.017g
H3BO3 ..................................................................0.01g
Na2MoO4 ·2H2O..........................................................0.01g
Preparation of Mineral Solution : Add nitrilotriacetic acid to
500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add
remaining components. Add distilled/deionized water to 1.0L. Mix
thoroughly.
Vitamin Solution:
Composition per liter:
Nicotinic acid.........................................................2.5mg
Thiamine·HCl .........................................................1.25mg
p-Aminobenzoic acid....................................................1.25mg
Pantothenic acid.......................................................0.62mg
Pyridoxine·HCl.........................................................6.2mg
Biotin.................................................................0.25mg
Preparation of Vitamin Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
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.
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 916.0mL. Adjust pH to 7.2. Gently heat and
bring to boiling. Continue boiling for a few minutes. Allow to cool to
room temperature under 80% N2 + 20% CO2 . Distribute into bottles
under 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution B:
Composition per 70.0mL:
NaHCO3 ................................................................3.5g
Preparation of Solution B:  Add NaHCO3 to distilled/deionized
water and bring volume to 70.0mL. Mix thoroughly. Filter sterilize.
Sparge with 80% N2 + 20% CO2 for 15 min.
Solution C:
Composition per 10.0mL:
L -Cysteine·HCl.........................................................0.3g
Preparation of Solution C: Add  L -cysteine·HCl to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Sparge with
100% N 2 for 3–4 min. Autoclave under 100% N2  for 15 min at 15 psi
pressure–121°C.
Solution D:
Composition per 10.0mL:
Na2S·9H2O...............................................................0.3g
Preparation of Solution D: Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 for 3–4 min. Autoclave under 100% N2  for 15 min at 15 psi pres-
sure–121°C.
Solution E:
Composition per 10.0mL:
Sodium ferulate........................................................1.5g
Preparation of Solution E: Add sodium ferulate to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize. Sparge with 100% N2 for 3–4 min.
Preparation of Medium: To 916.0mL of sterile solution A, add
70.0mL of sterile solution B, 10.0mL of sterile solution C, 10.0mL of
sterile solution D, and 10.0mL of sterile solution E. Mix thoroughly.
Anaerobically and aseptically distribute into sterile tubes or flasks.
Use:  For the cultivation and maintenance of Eubacterium callanderi.