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SP 4 Medium (DSMZ Medium 1076)

[所属分类:培养基配方] [发布时间:2021-8-19] [发布人:网站管理员2] [阅读次数:] [返回]

SP 4 Medium

(DSMZ Medium 1076)

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

Composition per 510.2mL:

Tryptone ....................................................................................... 5.0g

Peptone ........................................................................................ 3.3g

NaCl.............................................................................................. 0.5g

Beef extract................................................................................... 0.3g

Yeast extract.................................................................................. 0.3g

Beef heart, solids from infusion.................................................... 0.2g

Fetal bovine serum (inactivated at 56°C, 1 hr)........................90.0mL

CMRL 1066, 10X with glutamine...........................................25.0mL

Yeast extract solution...............................................................17.5mL

Yeastolate solution.....................................................................5.0mL

Glutamine solution.....................................................................1.7mL

Phenol Red solution...................................................................1.0mL

pH 7.4 ± 0.2 at 25°C

CMRL 1066, 10X with Glutamine:

Composition per liter:

NaCl.............................................................................................. 6.8g

NaHCO3........................................................................................ 2.2g

D-Glucose...................................................................................... 1.0g

KCl................................................................................................ 0.4g

L-Cysteine·HCl·H2O ................................................................... 0.26g

CaCl2, anhydrous.......................................................................... 0.2g

MgSO4·7H2O................................................................................ 0.2g

NaH2PO4·H2O............................................................................. 0.14g

L-Glutamine .................................................................................. 0.1g

Sodium acetate·3H2O................................................................ 0.083g

L-Glutamic acid......................................................................... 0.075g

L-Arginine·HCl............................................................................ 0.07g

L-Lysine·HCl ............................................................................... 0.07g

L-Leucine .................................................................................... 0.06g

Glycine........................................................................................ 0.05g

Ascorbic acid .............................................................................. 0.05g

L-Proline...................................................................................... 0.04g

L-Tyrosine ................................................................................... 0.04g

L-Aspartic acid ............................................................................ 0.03g

L-Threonine................................................................................. 0.03g

L-Alanine................................................................................... 0.025g

L-Phenylalanine......................................................................... 0.025g

L-Serine ..................................................................................... 0.025g

L-Valine ..................................................................................... 0.025g

L-Cystine ..................................................................................... 0.02g

L-Histidine·HCl·H2O................................................................... 0.02g

L-Isoleucine................................................................................. 0.02g

Phenol Red.................................................................................. 0.02g

L-Methionine............................................................................. 0.015g

Deoxyadenosine.......................................................................... 0.01g

Deoxycytidine............................................................................. 0.01g

Deoxyguanosine ......................................................................... 0.01g

Glutathione, reduced................................................................... 0.01g

Thymidine................................................................................... 0.01g

Hydroxy-L-proline ...................................................................... 0.01g

L-Tryptophan............................................................................... 0.01g

Nicotinamide adenine dinucleotide ........................................... 7.0mg

Tween™ 80................................................................................ 5.0mg

Sodium glucoronate·H2O........................................................... 4.2mg

Coenzyme A ..............................................................................2.5mg

Cocarboxylase............................................................................1.0mg

Flavin adenine dinucleotide .......................................................1.0mg

Nicotinamide adenine dinucleotide phosphate ..........................1.0mg

Uridine triphosphate ..................................................................1.0mg

Choline chloride.........................................................................0.5mg

Cholesterol .................................................................................0.2mg

5-Methyldeoxycytidine..............................................................0.1mg

Inositol .....................................................................................0.05mg

p-Aminobenzoic acid...............................................................0.05mg

Niacin.....................................................................................0.025mg

Niacinamide ...........................................................................0.025mg

Pyridoxine..............................................................................0.025mg

Pyridoxal·HCl ........................................................................0.025mg

Biotin .......................................................................................0.01mg

D-Calcium pantothenate ...........................................................0.01mg

Folic acid..................................................................................0.01mg

Riboflavin ................................................................................0.01mg

Thiamine·HCl ..........................................................................0.01mg

Preparation of CMRL 1066, 10X with Glutamine: Add com

ponents to distilled/deionized water and bring volume to 1.0L. Mix

thoroughly. Adjust pH to 7.2. Filter sterilize.

Yeast Extract Solution:

Composition per 10.0mL:

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

Preparation of Yeast Extract Solution: Add yesat extract to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Autoclave for 15 min at 15 psi pressure–121°C.

Yeastolate Solution:

Composition per 10.0mL:

Yeastolate...................................................................................... 2.0g

Preparation of Yeastolate Solution: Add yeastolate to distilled/

deionized water and bring volume to 10.0mL. Mix thoroughly. Auto

clave for 15 min at 15 psi pressure–121°C.

Phenol Red Solution:

Composition per 100.0mL:

Phenol Red.................................................................................... 1.0g

Preparation of Phenol Red Solution: Add 1.0g of Phenol Red to

distilled/deionized water and bring volume to 100.0mL. Mix

thoroughly. Adjust pH to 7.0. Filter sterilize.

Glutamine Solution:

Composition per 10.0mL:

L-Glutamine .................................................................................. 1.5g

Preparation of Glutamine Solution: Add 1.5g of L-glutamine to

distilled/deionized water and bring volume to 10.0mL. Mix

thoroughly. Filter sterilize.

Preparation of Medium: Add components, except fetal bovine se

rum, CMRL, yeast extract solution, yeastolate solution, Phenol Red so

lution, and glutamine solution, to distilled/deionized water and bring

volume to 375.0mL. Mix thoroughly. Adjust pH to 7.4. Gently heat and

bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool

to room temperature. Mix thoroughly. Aseptically add fetal bovine se

rum, CMRL, yeast extract solution, yeastolate solution, Phenol Red so

lution, and glutamine solution. Mix thoroughly.

Use: For the cultivation of Mycoplasma fermentans.

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SP 4 Medium (DSMZ Medium 1076)