Rhodobacter veldkampii Medium

[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

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

Composition per 127.0mL:

Solution 1.................................................................................76.2mL
Solution 2 + Solution 3............................................................44.8mL
Solution 4...................................................................................6.0mL
Solution 1:
Composition per 2.5L:
CaCl 2 .............................................................................................2.0g
Preparation of Solution 1: Add CaCl 2 to distilled/deionized water
and bring volume to 2.5L. Distribute in 80.0mL volumes into 127.0mL
screw-capped bottles. Autoclave for 15 min at 15 psi pressure–121°C.
Solution 2:
Composition per 100.0mL:
Sodium ascorbate..........................................................................2.4g
Sodium acetate..............................................................................1.0g
KC1...............................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
MgCl 2 ·6H 2 O..................................................................................0.8g
NH 4 Cl ...........................................................................................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:
Ethylenediamine tetraacetate (EDTA)..........................................1.5g
FeSO 4 ·7H 2 O..................................................................................0.2g
ZnSO 4 ·7H 2 O.................................................................................0.1g
MnCl 2 ·4H 2 O................................................................................0.02g
Modified Hoagland trace elements solution..............................6.0mL
Preparation of Heavy Metal Solution: Dissolve EDTA in 800.0mL
of distilled/deionized water. Add remaining components. Bring volume
to 1.0L with distilled/deionized water. Mix thoroughly.
Modified Hoagland Trace Elements Solution:
Composition  per 3.6L:
H 3 BO 3 ......................................................................................... 11.0g
MnCl 2 ·4H 2 O .................................................................................7.0g
AlCl 3 .............................................................................................1.0g
CoCl 2 ............................................................................................1.0g
CuCl 2 ............................................................................................1.0g
KI..................................................................................................1.0g
NiCl 2 .............................................................................................1.0g
ZnCl 2 .............................................................................................1.0g
BaCl 2 .............................................................................................0.5g
KBr ...............................................................................................0.5g
LiCl...............................................................................................0.5g
Na 2 MoO 4 ......................................................................................0.5g
SeCl 4 .............................................................................................0.5g
SnCl 2 ·2H 2 O...................................................................................0.5g
NaVO 3 ·H 2 O..................................................................................0.1g
Preparation of Modified Hoagland Trace Elements Solu-
tion: Prepare each component as a separate solution. Dissolve each
salt in approximately 100.0mL of distilled/deionized water. Adjust the
pH of each solution to below 7.0. Combine all the salt solutions and
bring the volume to 3.6L with distilled/deionized water. Adjust the pH
to 3–4. A yellow precipitate may form after mixing. After a few days,
it will turn into a fine white precipitate. Mix the solution thoroughly be-
fore using.
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.
Vitamin B 12  Solution:
Composition per 100.0mL:
Vitamin B 12  (cyanocobalamin) ..................................................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.
Solution 3:
Composition per 900.0mL:
NaHCO 3 ........................................................................................4.5g
Preparation of Solution 3:  Add NaHCO 3 to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Bubble 100%
CO 2 through the solution for 30 min. After CO 2 saturation of solution
3, add solution 2 and immediately filter the mixture through a Seitz fil-
ter (or a Millipore) using positive CO 2 pressure to push the liquid
through.
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 and bring volume to 200.0mL. Add a magnetic stir bar to the flask.
Autoclave for 15 min at 15 psi pressure–121°C. On a magnetic stirrer,
slowly add 2.0mL of sterile 2M H 2 SO 4 . This partially neutralizes the so-
lution. The solution should turn yellow. H 2 S gas will be liberated—neu-
tralization and distribution of the solution should be done as rapidly as
possible under adequate ventilation.
Preparation of Medium: To the 80.0mL of sterile solution 1 in
screw-capped bottles, add combined solutions 2 and 3 immediately af-
ter filtration and fill bottles to capacity. Mix thoroughly. Aseptically re-
move 6.0mL of the medium from the bottles and replace it with 6.0mL
of neutralized solution 4. Let stand for 24 hr. The medium should form
a fine white precipitate before using. To inoculate, remove 6.0mL of
the completed medium from the bottles and replace it with 6.0mL of
inoculum.
Use:  For the cultivation and maintenance of Rhodobacter veldkampii.