Medium SP 4

[所属分类:培养基配方] [发布时间:2021-5-20] [发布人:网站管理员2] [阅读次数:] [返回]

Medium SP 4

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

Composition per liter:

Pancreatic digest of casein.......................................................... 11.0g
Peptone .........................................................................................5.3g
Glucose.........................................................................................5.0g
NaCl..........................................................................................0.875g
Beef extract...............................................................................0.525g
Yeast extract..............................................................................0.525g
Beef heart, solids from infusion..................................................0.35g
Fetal bovine serum, heat inactivated .....................................170.0mL
Yeast extract solution.............................................................100.0mL
CMRL 1066, 10X solution......................................................50.0mL
Fresh yeast extract solution .....................................................35.0mL
Phenol Red solution.................................................................20.0mL
Penicillin solution....................................................................10.0mL
pH 7.6 ± 0.2 at 25°C
Yeast Extract Solution:
Composition per 100.0mL:
Yeast extract..................................................................................2.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
CMRL 1066, 10X Solution:
Composition per liter:
NaCl..............................................................................................6.8g
NaHCO 3 ........................................................................................2.2g
D- Glucose......................................................................................1.0g
KCl................................................................................................0.4g
L- Cysteine·HCl·H 2 O....................................................................0.26g
CaCl 2 , anhydrous..........................................................................0.2g
MgSO 4 ·7H 2 O................................................................................0.2g
NaH 2 PO 4 ·H 2 O.............................................................................0.14g
L- Glutamine...................................................................................0.1g
Sodium acetate·3H 2 O................................................................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·H 2 O...................................................................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·H 2 O...........................................................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, 10X medium is available as a premixed powder
from BD Diagnostics.
Preparation of CMRL 1066, 10X Solution: Add components 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. Filter sterilize.
Phenol Red Solution:
Composition per 100.0mL:
Phenol Red..................................................................................0.01g
Preparation of Phenol Red Solution: Add Phenol Red to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize.
Penicillin Solution:
Composition per 10.0mL:
Penicillin........................................................................... 1,000,000U
Preparation of Penicillin Solution: Add penicillin to distilled/de-
ionized water and bring volume to 10.0mL. Filter sterilize.
Preparation of Medium: Add components—except fetal bovine
serum, yeast extract solution, CMRL 1066, 10X solution, fresh yeast
extract solution, Phenol Red solution, and penicillin solution—to dis-
tilled/deionized water and bring volume to 615.0mL. Mix thoroughly.
Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to 45°–50°C. Aseptically add 170.0mL of sterile fe-
tal bovine serum, 100.0mL of sterile yeast extract solution, 50.0mL of
sterile CMRL 1066, 10X solution, 35.0mL of sterile fresh yeast extract
solution, 20.0mL of sterile Phenol Red solution, and 10.0mL of sterile
penicillin solution. Mix thoroughly. Aseptically distribute into sterile
tubes or flasks.
Use: For the isolation and cultivation of Spiroplasma species from
ticks.