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SP 4 Medium

[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

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

Composition per liter:

Base solution..........................................................................615.0mL

Fetal calf serum (inactivated at 56°C, 1 hr)...........................170.0mL

Yeast extract (2% solution)....................................................100.0mL

CMRL 1066, 10X with glutamine...........................................50.0mL

Fresh yeast extract solution .....................................................35.0mL

Phenol Red (0.1% solution).....................................................20.0mL

Penicillin solution ....................................................................10.0mL

pH 7.0–7.4 ± 0.2 at 25°C

Base Solution:

Composition per 615.0mL:

Pancreatic digest of casein.......................................................... 11.2g

Noble agar..................................................................................... 8.0g

Pancreatic digest of gelatin........................................................... 5.3g

Glucose ......................................................................................... 5.0g

NaCl.......................................................................................... 0.875g

Beef extract............................................................................... 0.525g

Yeast extract.............................................................................. 0.525g

Beef heart, solids from infusion.................................................. 0.35g

Preparation of Base Solution: Add components to distilled/deion

ized water and bring volume to 615.0mL. Mix thoroughly. Adjust pH

to 7.5. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi

pressure–121°C. Cool to 45°–50°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.

Fresh Yeast Extract Solution:

Composition per 100.0mL:

Baker’s yeast, live, pressed, starch-free,..................................... 25.0g

Preparation of Fresh Yeast Extract Solution: Add the live Bak

er’s yeast to 100.0mL of distilled/deionized water. Autoclave for 90

min at 15 psi pressure–121°C. Allow to stand. Remove supernatant so

lution. Adjust pH to 6.6–6.8.

Penicillin Solution:

Composition per 10.0mL:

Penicillin G ....................................................................... 1,000,000U

Preparation of Penicillin Solution: Add penicillin G to distilled/

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

sterilize.

Preparation of Medium: To 615.0mL of cooled sterile base solu

tion, aseptically add 170.0mL of sterile inactivated fetal calf serum,

100.0mL of sterile yeast extract, 50.0mL of sterile CMRL 1066, 10X

with glutamine, 35.0mL of sterile fresh yeast extract solution, 20.0mL

of Phenol Red solution, and 10.0mL of sterile penicillin solution. Mix

thoroughly. Aseptically distribute into sterile tubes. Allow tubes to

cool in a slanted position.

Use: For the cultivation of tick-derived Mycoplasma (Spiroplasma).

Used for the enhanced recovery of Mycoplasma pneumoniae, Myco

plasma alvi, and Mycoplasma hyopneumoniae.

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SP 4 Medium