Chlorobiaceae Medium 1

[所属分类:培养基配方] [发布时间:2020-4-20] [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

Chlorobiaceae Medium 1

培养基配方
Composition per 4990.0mL:
Solution 1.....................................................4.0L
O2 -free water..............................................860.0mL
NaHCO3 solution.............................................100.0mL
Na2S·9H2O solution..........................................20.0mL
Trace elements solution ......................................5.0mL
Vitamin B 12 solution.........................................5.0mL
pH 6.8 ± 0.2 at 25°C
Solution 1:
Composition  per 4.0L:
MgSO4 ·7H2O...................................................2.5g
KCl............................................................1.7g
KH2PO4 ........................................................1.7g
NH4Cl .........................................................1.7g
CaCl2 ·2H2O..................................................1.25g
Preparation of Solution 1:  Add components to distilled/deionized
water and bring volume to 4.0L. Mix thoroughly. Autoclave for 45 min
at 15 psi pressure–121°C. Cool to 25°C under 100% N2 . Saturate with
CO2 by stirring under 100% CO2 for 30 min.
O2 -Free Water:
Composition per 860.0mL:
H2O..........................................................860.0mL
Preparation of O2 -Free Water:  Autoclave H2O for 15 min at 15
psi pressure–121°C. Cool to 25°C under 100% N2 .
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ........................................................7.5g
Preparation of NaHCO3 Solution:  Add the NaHCO3 to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Gas
with 100% CO2 for 20 min. Filter sterilize with positive CO2 pressure.
Na2S·9H2O Solution:
Composition per 100.0mL:
Na2S·9H2O.........................................................10.0g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water. Mix thoroughly. Gas with 100% N2 for 15 min
in a screw-capped bottle. Tightly close cap. Autoclave for 15 min at 15
psi pressure–121°C. Cool to 25°C.
Trace Elements Solution:
Composition per liter:
FeCl2 ·4H2O......................................................1.5g
CoCl2 ·6H2O.....................................................0.19g
MnCl2 ·4H2O .....................................................0.1g
ZnCl2 ...........................................................0.07g
H3BO3 ...........................................................0.06g
NaMoO4 ·2H2O....................................................0.04g
CuCl2 ·2H2O.....................................................0.02g
NiCl2 ·6H20.....................................................0.02g
HCl(25% solution)................................................6.5mL
Preparation of Trace Elements Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Vitamin B 12 Solution:
Composition per 100.0mL:
Vitamin B 12 ....................................................2.0mg
Preparation of Vitamin B 12 Solution: Add vitamin B 12 to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Filter sterilize.
Preparation of Medium:  To 4.0L of sterile, CO2 -saturated solution
1, aseptically add the remaining components. Mix thoroughly. Adjust
pH to 6.8. Aseptically distribute into sterile 100.0mL bottles using pos-
itive pressure of 95% N2 + 5% CO2 . Completely fill bottles with medi-
um except for a pea-sized air bubble.
Use: For the isolation and cultivation of members of the Chloro-
biaceae.