Chromatium Medium (ATCC Medium 37)

[所属分类:培养基配方] [发布时间:2020-4-28] [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

Chromatium Medium
(ATCC Medium 37)

培养基配方
Composition per 127.0mL:
Solution 1............................................................76.2mL
Solution 2 + Solution 3...............................................44.8mL
Solution 4.............................................................6.0mL
Solution 1:
Composition per 2.5 L:
CaCl2 ..................................................................2.0g
Preparation of Solution 1: Add CaCl2 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
KC1.....................................................................1.0g
KH2PO4 .................................................................1.0g
MgCl2 ·6H2O ...........................................................0.8g
NH4Cl ..................................................................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
FeSO4 ·7H2O............................................................0.2g
ZnSO4 ·7H2O............................................................0.1g
MnCl2 ·4H2O ..........................................................0.02g
Modified Hoagland trace elements solution..............................6.0mL
Preparation of Heavy Metal Solution: Dissolve EDTA in ap-
proximately 800.0mL of distilled/deionized water. Add remaining
components and bring volume to 1.0L. Mix thoroughly.
Modified Hoagland Trace Elements Solution:
Composition  per 3.6L:
H3BO3 ................................................................11.0g
MnCl2 ·4H2O ..........................................................7.0g
AlCl3 .................................................................1.0g
CoCl2 .................................................................1.0g
CuCl2 .................................................................1.0g
KI.....................................................................1.0g
NiCl2 .................................................................1.0g
ZnCl2 .................................................................1.0g
BaCl2 .................................................................0.5g
KBr ...................................................................0.5g
LiCl...................................................................0.5g
Na2MoO4 ...............................................................0.5g
SeCl4 .................................................................0.5g
SnCl2 ·2H2O...........................................................0.5g
NaVO3 ·H2O............................................................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 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.
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.
Solution 3:
Composition per 900.0mL:
NaHCO3 .............................................................4.5g
Preparation of Solution 3:  Add NaHCO3 to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Bubble 100%
CO2 through the solution for 30 min. After CO2 saturation of solution
3, add solution 2 and immediately filter the mixture through a Seitz fil-
ter (or a Millipore) using positive CO2 pressure to push the liquid
through.
Solution 4:
Composition per 200.0mL:
Na2S·9H2O..........................................................3.0g
Preparation of Solution 4: Add Na2S·9H2O to distilled/deionized wa-
ter and bring volume to 200.0mL. Add a magnetic stir bar to the flask. Au-
toclave for 15 min at 15 psi pressure–121°C. On a magnetic stirrer, slowly
add 2.0mL of sterile 2MH2SO4 . This partially neutralizes the solution. The
solution should turn yellow. H2S gas will be liberated. Neutralization and
distribution of the solution should be done as rapidly as possible under ad-
equate 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 Chromatium tepidum.