C 3N 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 3N Spiroplasma Medium

培养基配方
Composition per 100.0mL:
Sucrose........................................................12.0g
Phenol Red....................................................10.0mg
PPLO broth without Crystal Violet.............................50.0mL
Horse serum...................................................20.0mL
Fresh yeast extract solution ..................................5.0mL
CMRL-1066 medium...............................................0.5mL
pH 7.5 ± 0.2 at 25°C
PPLO Broth without Crystal Violet:
Composition per 500.0mL:
Beef heart, infusion from....................................11.52g
Peptone .....................................................2.32g
NaCl.........................................................1.15g
Source: PPLO broth without Crystal Violet is available as a premixed
powder from BD Diagnostic Systems.
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 75.0mL. Adjust pH to 7.5. Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically add 20.0mL of sterile horse se-
rum, 5.0mL of sterile yeast extract, and 0.5mL of sterile CMRL medi-
um. Distribute into sterile tubes or flasks.
Use:  For the cultivation and maintenance of Spiroplasma kunkelii.