[所屬分類:培養(yǎng)基配方] [發(fā)布時(shí)間:2020-7-16] [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

Filobacillus milosensis Medium
(DSMZ Medium 607a)

培養(yǎng)基配方
Composition per liter:
NaCl....................................................................100.0g
Peptone...................................................................75g
Yeast extract............................................................0.75g
Glucose .................................................................0.75g
Artificial sea water....................................................250.0mL
Tris/HCl (0.1M, pH 7.5) .................................................50.0mL
Hutner's basal salts solution............................................20.0mL
Vitamin solution.........................................................10.0mL
pH 7.2 ± 0.2 at 25°C
Hutner’s Basal Salts Solution:
Composition per liter:
MgSO4 ·7H2O.............................................................29.7g
Nitrilotriacetic acid....................................................10.0g
CaCl2 ·2H2O............................................................3.335g
FeSO4 ·7H2O............................................................99.0mg
(NH4)6 MoO7O24 ·4H2O ..................................................9.25mg
"Metals 44" ............................................................50.0mL
"Metals 44":
Composition per 100.0mL:
ZnSO4 ·7H2O............................................................1.095g
FeSO4 ·7H2O..............................................................0.5g
Sodium EDTA..............................................................0.25g
MnSO4 ·H2O .............................................................0.154g
CuSO4 ·5H2O.............................................................39.2mg
Co(NO3)2 ·6H2O..........................................................24.8mg
Na2B4O7 ·10H2O..........................................................17.7mg
Preparation of “Metals 44”: Add sodium EDTA to distilled/de-
ionized water and bring volume to 90.0mL. Mix thoroughly. Add a few
drops of concentrated H2SO4 to retard precipitation of heavy metal
ions. Add remaining components. Mix thoroughly. Bring volume to
100.0mL with distilled/deionized water.
Preparation of Hutner’s Basal Salts Solution:  Add nitrilotria-
cetic 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. Adjust pH to 6.8.
Artificial Sea Water:
Composition per liter:
NaCli.....................................................................23.477g
MgCl2 ·6H2O ..............................................................4.981g
Na2SO4 ....................................................................3.917g
CaCl2 .....................................................................1.12g
KCl.......................................................................664.0mg
NaHCO3 ...................................................................192.0mg
H3BO3 ....................................................................26.0mg
SrCl2 ....................................................................24.0mg
KBr .......................................................................6.0mg
NaF .......................................................................3.0mg
Preparation of Artificial Sea Water:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
Vitamin Solution:
Composition per liter:
Riboflavin................................................................5.0mg
Nicotinamide..............................................................5.0mg
Thiamine-HCl·2H2O........................................................5.0mg
Ca-pantothenate...........................................................5.0mg
Biotin....................................................................2.0mg
Folic acid ...............................................................2.0mg
Vitamin B 12 .............................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Add components, except artificial sea water
and vitamin solution, to distilled/deionized water and bring volume to
740.0mL. Mix thoroughly. Adjust pH to 7.2. Autoclave for 15 min at 15
psi pressure–121°C. Cool to room temperature. Aseptically add 250.0mL
artificial sea water and 10.0mL vitamin solution. Mix thoroughly. Asepti-
cally and anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Filobacillus milosensis.