C 3G Spiroplasma Medium

[所属分类:培养基配方] [发布时间:2020-3-25] [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

C 3G Spiroplasma Medium

培养基配方
Composition per liter:
Sucrose.......................................................100.0g
Phenol Red....................................................10.0mg
PPLO broth without Crystal Violet............................500.0mL
Horse serum..................................................150.0mL
Fresh yeast extract solution .................................50.0mL
CMRL-1066 medium...............................................5.0mL
pH 7.5 ± 0.2 at 25°C
Source: PPLO broth without Crystal Violet is available as a premixed
powder from BD Diagnostic Systems.
PPLO Broth without Crystal Violet:
Composition per 500.0mL:
Beef heart, infusion from.......................................11.52g
Peptone .........................................................2.32g
NaCl.............................................................1.15g
Preparation of PPLO Broth without Crystal Violet:  Add
components to distilled/deionized water and bring volume to 500.0mL.
Mix thoroughly.
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. Filter sterilize.
CMRL-1066 Medium:
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
Source:  CMRL-1066 medium is available as a premixed powder
from BD Diagnostics.
Preparation of CMRL-1066 Medium: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Ad-
just pH to 7.2. Filter sterilize.
Preparation of Medium: Add components—except horse serum,
fresh yeast extract, and CMRL medium—to distilled/deionized water
and bring volume to 795.0mL. Adjust pH to 7.5. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically add 150.0mL of sterile horse se-
rum, 50.0mL of sterile fresh yeast extract solution, and 5.0mL of sterile
CMRL medium. Distribute into sterile tubes or flasks.
Use:  For the cultivation and maintenance of Spiroplasma species.