Kelly Medium, Selective Modified

[所属分类:培养基配方] [发布时间:2021-4-22] [发布人:李佳颐] [阅读次数:] [返回]

Kelly Medium, Selective Modified

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

Composition per 1270.0mL:

Bovine serum albumin fraction V...............................................50.0g
HEPES buffer (N-2-hydroxyethylpiperazine-
N-2-ethanesulfonic acid)........................................................6.0g
Glucose.........................................................................................5.0g
Neopeptone...................................................................................5.0g
NaHCO 3 ........................................................................................2.2g
Sodium pyruvate...........................................................................0.8g
Sodium citrate...............................................................................0.7g
N-Acetylglucosamine ...................................................................0.4g
Kanamycin.................................................................................8.0mg
5-Fluorouracil............................................................................2.3mg
Gelatin solution......................................................................200.0mL
CMRL-1066 medium with glutamine, 10X...........................100.0mL
Rabbit serum, partially hemolyzed..........................................70.0mL
pH 7.7 ± 0.2 at 25°C
Gelatin Solution:
Composition per 200.0mL:
Gelatin.........................................................................................14.0g
Preparation of Gelatin Solution: Add gelatin to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly. Gently heat and
bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 50°C.
CMRL-1066 Medium with Glutamine, 10X:
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 glucuronate·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
pH 7.2 ± 0.2 at 25°C
Source:  This solution is available as a premixed powder from BD Di-

agnostics.

Preparation of CMRL-1066 Medium with Glutamine, 10X:

Add components to distilled/deionized water and bring volume to
1.0L. Mix thoroughly. Adjust pH to 7.2. Filter sterilize.
Preparation of Medium: Add components, except gelatin solution,
partially hemolyzed rabbit serum, kanamycin, and 5-fluorouracil, to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Bring pH to 7.6 with 5N NaOH. Filter sterilize. Aseptically add
200.0mL of sterile gelatin solution, 70.0mL of partially hemolyzed
rabbit serum, 8.0mg of kanamycin, and 230.0mg of 5-fluorouracil. Mix
thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the isolation of Borrelia burgdorferi.