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培养基配方

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Thermotoga elfii Medium

[所属分类:培养基配方] [发布时间:2021-9-8] [发布人:网站管理员2] [阅读次数:] [返回]

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

Composition per liter:

NaCl ............................................................................................ 10.0g

Pancreatic digest of casein............................................................ 2.0g

Yeast extract.................................................................................. 2.0g

NH4Cl ........................................................................................... 1.0g

K2HPO4......................................................................................... 0.3g

KH2PO4......................................................................................... 0.3g

MgCl2·6H2O.................................................................................. 0.2g

CaCl2·2H2O................................................................................... 0.1g

KCl................................................................................................ 0.1g

L-Cysteine·HCl·H2O ..................................................................... 0.5g

Sodium acetate.............................................................................. 0.5g

Resazurin ...................................................................................1.0mg

Na2CO3 solution ......................................................................20.0mL

Na2S·9H2O solution .................................................................20.0mL

Modified Wolfe’s mineral solution ..........................................10.0mL

pH 8.0 ± 0.2 at 25°C

NaHCO3 Solution:

Composition per 20.0mL:

NaHCO3 ........................................................................................ 2.0g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge

with 80% N2 + 20% CO2. Autoclave for 15 min at 15 psi pressure–

121°C.

Na2S·9H2O Solution:

Composition per 20.0mL:

Na2S·9H2O.................................................................................... 0.4g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.

Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C.

Before use, neutralize to pH 7.0 with sterile HCl.

Modified Wolfe’s Mineral Solution:

Composition per liter:

MgSO4·7H2O ................................................................................ 3.0g

Nitrilotriacetic acid ....................................................................... 1.5g

NaCl .............................................................................................. 1.0g

MnSO4·H2O .................................................................................. 0.5g

CaCl2............................................................................................. 0.1g

CoCl2·6H2O .................................................................................. 0.1g

FeSO4·7H2O.................................................................................. 0.1g

ZnSO4·7H2O ................................................................................. 0.1g

AlK(SO4)2·12H2O....................................................................... 0.01g

CuSO4·5H2O ............................................................................... 0.01g

H3BO3 ......................................................................................... 0.01g

Na2MoO4·2H2O .......................................................................... 0.01g

Na2SeO3 ...................................................................................... 0.01g

NaWO4·2H2O ............................................................................. 0.01g

NiC12·6H2O................................................................................ 0.01g

Preparation of Modified Wolfe’s Mineral Solution: Add

nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH

to 6.5 with KOH. Add remaining components one at a time. Add dis

tilled/deionized water to 1.0L. Adjust pH to 6.8.

Preparation of Medium: Prepare and dispense medium under

100% N2. Add components, except Na2CO3 solution and Na2S·9H2O

solution, to distilled/deionized water and bring volume to 960.0mL.

Mix thoroughly. Adjust pH to 8.0 with 10M KOH. Gently heat and

bring to boiling. Cool to room temperature while sparging with 100%

N2. Anaerobically dispense into tubes in 5.0mL aliquots under an at

mosphere of 80% N2 + 20% CO2. Autoclave for 45 min at 6 psi pres

sure–110°C. Just prior to use, aseptically and anaerobically add 0.1mL

of sterile Na2CO3 solution and 0.1mL of sterile Na2S·9H2O solution to

each tube.

Use: For the cultivation of Thermotoga elfii.

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Thermotoga elfii Medium