Rhodobacter veldkampii Medium (DSMZ Medium 867)

[所属分类:培养基配方] [发布时间:2021-8-4] [发布人:网站管理员2] [阅读次数:] [返回]

Rhodobacter veldkampii Medium
(DSMZ Medium 867)

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

Composition per 2780.0mL:

Solution 1.............................................................................1540.0mL
Solution 3.............................................................................1000.0mL
Solution 4...............................................................................120.0mL
Solution 5...............................................................................120.0mL
pH 4.0 ± 0.1 at 25°C
Solution 1:
Composition per 2500.0mL:
CaCl 2 .............................................................................................2.0g
Preparation of Solution 1: Add CaCl 2 to distilled/deionized water
and bring volume to 2.5L. Mix thoroughly.
Solution 3:
Composition per liter:
NaHCO 3 ........................................................................................4.5g
Solution 2...............................................................................100.0mL
Preparation of Solution 3:  Add NaHCO 3 to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Sparge with gas-
eous CO 2 for at least 30 min. Add 100.0mL solution 2. Immediately fil-
ter sterilize using CO 2 pressure to push the liquid through (no suction).
Solution 2:
Composition per 100.0mL:
Sodium ascorbate..........................................................................2.4g
KH 2 PO 4 .........................................................................................1.0g
KCl................................................................................................1.0g
NH 4 Cl ...........................................................................................0.8g
MgCl 2 ·6H 2 O..................................................................................0.8g
Heavy metal solution...............................................................50.0mL
Vitamin solution.......................................................................15.0mL
Vitamin B 12 solution..................................................................3.0mL
Preparation of Solution 2:  Add components to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly.
Heavy Metal Solution:
Composition per liter:
EDTA..........................................................................................1.50g
FeSO 4 ·7H 2 O..................................................................................0.2g
ZnSO 4 ·7H 2 O.................................................................................0.1g
MnCl 2 ·7H 2 O ...............................................................................0.02g
Modified Hoagland trace elements solution..............................6.0mL
Preparation of Heavy Metal Solution:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Modified Hoagland Trace Elements Solution:
Composition per 3.6L:
H 3 BO 3 ......................................................................................... 11.0g
MnCl 2 ·4H 2 O .................................................................................7.0g
ZnCl 2 .............................................................................................1.0g
CuCl 2 ............................................................................................1.0g
NiCl 2 .............................................................................................1.0g
CoCl 2 ............................................................................................1.0g
AlCl 3 .............................................................................................1.0g
KI..................................................................................................1.0g
KBr ...............................................................................................0.5g
LiCl...............................................................................................0.5g
SnCl 2 ·2H 2 O...................................................................................0.5g
BaCl 2 ............................................................................................0.5g
Na 2 MoO 4 ......................................................................................0.5g
Na 2 SeO 3 ........................................................................................0.5g
NaVO 3 ·H 2 O..................................................................................0.1g
Preparation of Modified Hoagland Trace Elements Solu-
tion:  Add components sequentially to distilled/deionized water and
bring final volume to 3.6L. Mix thoroughly after adding each compo-
nent until dissolved. Adjust pH to just below 7.0. Adjust the final pH
to 3–4. The flaky yellow precipitate which is formed after mixing
transforms after standing for one or a few days into a very fine white
precipitate. Mix thoroughly before use.
Vitamin B 12 Solution:
Composition per 3.0mL:
Vitamin B 12 (cyanocobalamine)................................................2.0mg
Preparation of Vitamin B 12 Solution:  Add vitamin B 12 to dis-
tilled/deionized water and bring volume to 3.0mL. Mix thoroughly.
Vitamin Solution:
Composition per 100.0mL:
Pyridoxamine·2HCl...................................................................5.0mg
Nicotinic acid.............................................................................2.0mg
Thiamine....................................................................................1.0mg
Pantothenic acid.........................................................................0.5mg
Biotin.........................................................................................0.2mg
p-Aminobenzoic acid.................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 100% CO 2  for 30 min.
Solution 4:
Composition per 200.0mL:
Na 2 S·9H 2 O....................................................................................3.0g
Preparation of Solution 4: Add Na 2 S·9H 2 O to distilled/deionized
water in a flask with a magnetic stirrer and bring volume to 200.0mL.
Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pres-
sure–121°C. Cool to room temperature. Partially neutralize the steril-
ized solution by adding, on a magnetic stirrer, drop by drop, 2.0mL
sterile 2M H 2 SO 4 .
Solution 5:
Composition per 200.0mL:
Na-acetate .....................................................................................6.0g
Preparation of Solution 5:  Add Na-acetate to distilled/deionized
water and bring volume to 200.0mL. Mix thoroughly. Sparge with
100% N 2 for 5 min. Autoclave under 100% N 2 for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Preparation of Medium: Distribute solution 1 in 77.0mL amounts
into 20 127mL screw-capped Boston round bottles. Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature. Aseptically
add 50.0mL sterile solution 3 to each of the 20 127mL bottles contain-
ing 77.0mL of sterile solution 1 so that the solution completely fills the
bottle. Mix thoroughly. Remove 6.0mL of the medium from the com-
pletely filled bottles. Add 6.0mL of neutralized solution 4 so that the
bottles are again completely filled. Mix thoroughly. Remove 6.0mL of
the medium from the completely filled bottles. Add 6.0mL of solution
5 so that the bottles are again completely filled. Mix thoroughly. Adjust
pH to 4.0. Allow the bottles to stand overnight to develop a hazy, white
precipitate before inoculating. Mix the solution thoroughly before use.
To inoculate, remove 6.0mL of completed medium and replace it with
an equal volume of inoculum. Grow cultures under tungsten light.
Use:  For the cultivation of Rhodobacter veldkampii.