Bryant-Robinson Medium

[所属分类:培养基配方] [发布时间:2020-3-16] [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

Bryant-Robinson Medium

培养基配方
Composition per 1010.0mL:
Glucose, cellobiose, or maltose...............................5.0g
L- Methionine.................................................0.08g
Mineral solution.............................................50.0mL
Na2CO3 solution .............................................50.0mL
Hemin solution...............................................10.0mL
L- Cysteine·HCl–Na2S solution..............................10.0mL
Vitamin solution.............................................5.0mL
VFA solution..................................................4.5mL
Resazurin ....................................................1.0mL
pH 6.5 ± 0.2 at 25°C
Mineral Solution:
Composition per liter:
KH2PO4 .........................................................18.0g
NaCl............................................................18.0g
(NH4)2SO4 ......................................................8.0g
CaCl2 ·6H2O ...................................................0.53g
MgCl2 ·6H2O ...................................................0.4g
CoCl2 ·6H2O ...................................................0.2g
MnCl2 ·4H2O....................................................0.2g
FeSO4 ·7H2O....................................................0.08g
Preparation of Mineral Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Na2CO3 Solution:
Composition per 100.0mL:
Na2CO3 .............................................................8.0g
Preparation of Na 2 CO 3 Solution: Add Na 2 CO 3 to O 2 -free dis-
tilled/deionized water. Mix thoroughly. Gas with 100% CO 2 for 15
min. Autoclave for 15 min at 15 psi pressure–121°C.
Hemin Solution:
Composition per 100.0mL:
Hemin.............................................................0.01g
NaOH (0.002% solution)..........................................100.0mL
Preparation of Hemin Solution: Add hemin to 100.0mL of NaOH
solution. Mix thoroughly.
L -Cysteine·HCl–Na 2 S Solution:
Composition per 100.0mL:
L- Cysteine·HCl..................................................2.5g
Na2S·9H2O........................................................2.5g
Preparation of  L -Cysteine·HCl–Na 2 S Solution: Add  L -
cysteine·HCl to distilled/deionized water and bring volume to 80.0mL.
Mix thoroughly. Adjust pH to 11 with NaOH. Add Na2S·9H2O. Mix
thoroughly. Bring volume to 100.0mL with distilled/deionized water.
Gently heat and bring to boiling under 100% N2 . Cool to 25°C under
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin Solution:
Composition per 100.0mL:
Calcium pantothenate ......................................0.02g
Nicotinamide...............................................0.02g
Pyridoxine·HCl............................................0.02g
Riboflavin ................................................0.02g
Thiamine·HCl .............................................0.02g
p-Aminobenzoic acid........................................1.0mg
Biotin ....................................................0.25mg
Folic acid.................................................0.25mg
Vitamin B 12 ..............................................0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
VFA (Volatile Fatty Acid) Solution:
Composition per liter:
Acetic acid .....................................................36.0mL
DL- α-Methylbutyric acid........................................2.0mL
Isovaleric acid..................................................2.0mL
n-Valeric acid...................................................2.0mL
Isobutyric acid..................................................1.8mL
Preparation of VFA Solution: Add components to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Add components, except  L -cysteine·HCl–
Na2S solution, to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly. Gently heat and bring to boiling. Continue boiling until resa-
zurin turns colorless, indicating reduction. Anaerobically distribute into
tubes in 10.0mL volumes. Cap with butyl rubber stoppers. Place tubes in a
press. Autoclave for 15 min at 15 psi pressure–121°C. Immediately prior
to inoculation, aseptically and anaerobically add 0.1mL of  L -
cysteine·HCl–Na2S solution per tube.
Use: For the cultivation of Bacteroides species from rumens.