Selenate Reducer Medium (DSMZ Medium 1049)

[所属分类:培养基配方] [发布时间:2021-8-13] [发布人:网站管理员2] [阅读次数:] [返回]

Selenate Reducer Medium
(DSMZ Medium 1049)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1004mL:

Solution A..............................................................................940.0mL
Solution B..................................................................................9.4mL
Solution C................................................................................23.5mL
Solution D................................................................................12.2mL
Solution E ................................................................................0.94mL
Solution F (vitamin solution).....................................................4.7mL
Solution G (trace elements solution SL-10) ..............................9.4mL
Solution H..................................................................................4.7mL
pH 6.9 ± 0.2 at 25°C
Solution A :
Composition per 940.0mL:
NaCl ...........................................................................................23.0g
MgCl 2 ·6H 2 O..................................................................................3.0g
KCl .............................................................................................1.30g
DL -Na-lactate .............................................................................1.12g
NH4Cl ..........................................................................................0.5g
KH2PO4 .......................................................................................0.2g
CaCl 2 ·2H 2 O...................................................................................0.1g
Resazurin ..................................................................................0.5mg
Preparation of Solution A:  Add components to distilled/deionized
water and bring volume to 940.0mL. Mix thoroughly. Distribute to an-
aerobic cultivation vessels. Gently heat and bring to boiling. Cool to
room temperature while sparging with a gas mixture of 80% N 2 + 20%
CO 2 . Seal vessels. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to room temperature.
Solution B :
Composition per 10.0mL:
Na 2 SeO 4 ....................................................................................0.019g
Preparation of Solution B:  Add Na 2 SeO 4 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gently heat and
bring to boiling. Cool to room temperature while sparging with a gas
mixture of 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to room temperature.
Solution C :
Composition per 10.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of Solution C:  Add NaHCO 3 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gently heat and
bring to boiling. Cool to room temperature while sparging with a gas
mixture of 80% H 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to room temperature.
Solution D :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Solution D: Add Na 2 S·9H 2 O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Autoclave under
100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room tempera-
ture.
Solution E:
Composition per 10.0mL:
Na 2 WO 4 .....................................................................................8.0mg
Na 2 S 2 O 3 .....................................................................................3.0mg
Preparation of Solution E:  Add components to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N 2 . Filter sterilize.
Solution F (Vitamin Solution):
Composition per liter:
Folic acid ....................................................................................20.0g
α-Lipoic acid...........................................................................50.0mg
p-Aminobenzoic acid...............................................................50.0mg
Pantothenic acid.......................................................................50.0mg
Riboflavin................................................................................50.0mg
Thamine·HCl ...........................................................................50.0mg
Vitamin B 12 ..............................................................................50.0mg
Nicotine amide.........................................................................25.0mg
Biotin.......................................................................................20.0mg
Nicotinic acid...........................................................................20.0mg
Pyridoxamine·HCl...................................................................10.0mg
Preparation of Solution F (Vitamin Solution) : Add compo-
nents to distilled/deionized water and bring volume to 1.0L. Sparge
with 100% N 2 . Mix thoroughly. Stir for 2 hr. Filter sterilize.
Solution G (Trace Elements Solution SL-10):
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Solution G (Trace Elements Solution SL-10):
Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Dis-
solve by adjusting pH to 6.5 with KOH. Add remaining components.
Add distilled/deionized water to 1.0L. Mix thoroughly. Adjust pH to
7.0. Sparge with 100% N 2 . Filter sterilize.
Solution H:
Composition per 10.0mL:
1,4-Naphthaquinone ..................................................................0.4mg
Hemin .......................................................................................0.1mg
Preparation of Solution H:  Add components to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N 2 . Filter sterilize.
Preparation of Medium:  Add the following to 10.0mL of solution
A: 0.1mL solution B, 0.25mL solution C, 0.13mL solution D, 0.01mL
solution E, 0.05mL solution F, 0.1mL solution G, and 0.05mL solution
H.
Use: For the cultivation of Sedimenticola selenatireducens.