Desulfomonile tiedjei Medium

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfomonile tiedjei Medium

培养基配方
Composition per liter:
NaHCO3 ....................................................................3.0g
PIPES (piperazine-N,N′-bis-2-ethanesulfonic acid) buffer...................1.5g
Na2SO4 ...................................................................1.42g
Yeast extract..............................................................1.0g
Mineral solution..........................................................20.0mL
Trace metal solution......................................................10.0mL
Na2S2O4 solution .........................................................10.0mL
Vitamin solution..........................................................10.0mL
Sodium pyruvate solution..................................................10.0mL
Resazurin (0.1% solution) .................................................1.0mL
pH 7.3 ± 0.2 at 25°C
Mineral Solution:
Composition per liter:
NH4Cl ......................................................................50.0g
NaCl........................................................................40.0g
MgCl2 ·6H2O ................................................................8.3g
KCl..........................................................................5.0g
KH2PO4 ......................................................................5.0g
CaCl2 ·2H2O ................................................................1.0g
Preparation of Mineral Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Trace Metal Solution:
Composition per liter:
Nitrilotriacetic acid.........................................................2.0g
MnSO4· H2O...................................................................1.0g
Fe(NH4)2(SO4)2 ·6H2O.........................................................0.8g
CoCl2 ·6H2O..................................................................0.2g
ZnSO4 ·7H2O..................................................................0.2g
CuCl2 ·2H2O.................................................................0.02g
Na2MoO4 ·H2O................................................................0.02g
Na2SeO4 .....................................................................0.02g
Na2WO4 ......................................................................0.02g
NiCl2 ·6H2O.................................................................0.02g
Preparation of Trace Metal Solution: Add nitrilotriacetic acid to
500.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5
with KOH. Add remaining components. Add distilled/deionized water to
1.0L.
Vitamin Solution:
Composition per liter:
Nicotinamide..................................................................0.05g
1,4-Naphthoquinone ...........................................................0.02g
p-Aminobenzoic acid...........................................................5.0mg
Biotin........................................................................5.0mg
Calcium pantothenate .........................................................5.0mg
Cyanocobalamin................................................................5.0mg
Folic acid ...................................................................5.0mg
Hemin ........................................................................5.0mg
Pyridoxine·HCl...............................................................5.0mg
Riboflavin....................................................................5.0mg
Thioctic acid.................................................................5.0mg
NaOH (0.1N solution)..........................................................5.0mL
Preparation of Vitamin Solution:  Add thioctic acid, 1,4-naphtho-
quinone, and hemin to 5.0mL of 0.1N NaOH solution. Mix thoroughly.
Bring volume to 1.0L with distilled/deionized water. Add remaining
components. Mix thoroughly.
Na2S2O4 Solution:
Composition per 10.0mL:
Na2S2O4 .....................................................................0.087g
Preparation of Na2S2O4 Solution: Add Na2S2O4 to distilled/de-
ionized water and bring volume to 10.0mL. Filter sterilize. Prepare
freshly.
Sodium Pyruvate Solution:
Composition per 10.0mL:
Sodium pyruvate................................................................4.4g
Preparation of Sodium Pyruvate Solution: Add sodium pyru-
vate to distilled/deionized water and bring volume to 10.0mL. Filter
sterilize.
Preparation of Medium: Add PIPES buffer, Na2SO4 , yeast extract,
mineral solution, and trace metal solution to distilled/deionized water
and bring volume to 970.0mL. Mix thoroughly. Adjust pH to 7.3 with
HCl. Add NaHCO3 and resazurin. Gently heat and bring to boiling un-
der 80% N2 + 20% CO2 . Replace headspace with 2 atm pressure of the
same gas phase. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 25°C. Anaerobically and aseptically add sterile vitamin solution, so-
dium pyruvate solution, and Na2S2O4 solution. Mix thoroughly.
Use: For the cultivation and maintenance of Desulfomonile tiedjei.