Diazotrophic Medium (RBA)

[所属分类:培养基配方] [发布时间:2020-6-16] [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

Diazotrophic Medium
(RBA)

培养基配方
Composition per 1008.0mL:
Solution A...............................................................903.0mL
Solution B................................................................50.0mL
Solution C................................................................50.0mL
Solution D.................................................................5.0mL
pH 7.3 ± 0.2 at 25°C
Solution A:
Composition per 903.0mL:
Agar .....................................................................15.0g
K2HPO4 ....................................................................0.9g
CaCl2 ·2H2O...............................................................0.1g
KH2PO4 ....................................................................0.1g
MgSO4 ·7H2O...............................................................0.1g
NaCl.......................................................................0.1g
FeSO4 ·7H2O..............................................................0.01g
MnSO4 ·H2O...............................................................5.0mg
NaVO3 ·2H2O .............................................................5.0mg
Na2MoO4 ·2H2O ...........................................................0.5mg
Trace elements solution SL-6 .............................................3.0mL
Trace Elements Solution SL-6:
Composition per liter:
MnCl2 ·4H2O...............................................................0.5g
H3BO3 .....................................................................0.3g
CoCl2 ·6H2O ..............................................................0.2g
ZnSO4 ·7H2O...............................................................0.1g
Na2MoO4 ·2H2O............................................................0.03g
NiCl2 ·6H2O..............................................................0.02g
CuCl2 ·2H2O..............................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 7.3. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Solution A : Add components to distilled/deionized
water and bring volume to 903.0mL. Mix thoroughly. Gently heat and
bring to boiling. Adjust pH to 7.3. Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to 50°–55°C. Pour into sterile Petri dishes or distrib-
ute into sterile tubes.
Solution B:
Composition per 50.0mL:
DL -Malate..................................................................2.0g
Disodium succinate..........................................................1.0g
Yeast extract..............................................................0.05g
Preparation of Solution B : Add components to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Adjust pH to 7.3.
Autoclave for 15 min at 15 psi pressure–121°C.
Solution C:
Composition per 50.0mL:
D -Mannitol...............................................................2.0g
D -Glucose................................................................2.0g
Sodium pyruvate...........................................................1.0g
Preparation of Solution C : Add components to distilled/deionized
water and bring volume to 50.0mL. Mix thoroughly. Adjust pH to 7.3.
Filter sterilize.
Solution D:
Composition per liter:
Pyridoxine·HCl............................................................62.5g
Nicotinic acid............................................................25.0mg
p-Aminobenzoic acid.......................................................12.5mg
Thiamine·HCl ............................................................12.5mg
Calcium  DL -pantothenate..................................................6.5mg
Biotin.....................................................................2.5mg
Preparation of Solution D:  Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.5. Gas
under 100% N2 . Filter sterilize.
Preparation of Medium:  To 903.0mL of sterile solution A, aseptically
add 50.0mL of sterile solution B, 50.0mL of sterile solution C, and 5.0mL
of sterile solution D. Mix thoroughly. Final pH should be 7.3. Pour into
sterile Petri dishes or distribute into sterile tubes.
Use:  For the cultivation and maintenance of Arthrobacter species,
Azomonas species, Azorhizophilus paspali, and Azotobacter species.