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Kelly Medium, Nonselective 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, Nonselective Modified

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

Composition per 1430.0mL:

HEPES buffer (N-2-hydroxyethylpiperazine-
N-2-ethanesulfonic acid)........................................................6.0g
Proteose peptone No. 2.................................................................5.0g
D- Glucose......................................................................................3.0g
NaHCO 3 ........................................................................................2.2g
Pancreatic digest of casein............................................................1.0g
Yeast, autolyzed............................................................................1.0g
Sodium pyruvate...........................................................................0.8g
Sodium citrate...............................................................................0.7g
N-Acetylglucosamine ...................................................................0.4g
MgCl 2 ·6H 2 O .................................................................................0.3g
Gelatin solution......................................................................200.0mL
Bovine serum albumin solution.............................................143.0mL
CMRL-1066 medium with glutamine, 10X...........................100.0mL
Rabbit serum, heat inactivated.................................................86.0mL
Hemin solution...........................................................................1.0mL
pH 7.2 ± 0.2 at 25°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.

Gelatin Solution:
Composition per 200.0mL:
Gelatin.........................................................................................14.0g
Preparation of Gelatin Solution: Add gelatin to distilled/deion-
ized water and bring volume to 200.0mL. Mix thoroughly. Gently heat
and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 50°C.
Hemin Solution:
Composition per 100.0mL:
Hemin ...........................................................................................1.0g
NaOH (1N solution).................................................................20.0mL
Preparation of Hemin Solution:  Add hemin to 20.0mL of 1N
NaOH solution. Mix thoroughly. Bring volume to 100.0mL with dis-
tilled/deionized water.
Bovine Serum Albumin Solution:
Composition per 200.0mL:
Bovine serum albumin................................................................70.0g
Preparation of Bovine Serum Albumin Solution:  Add bovine
serum albumin to distilled/deionized water and bring volume to
200.0mL. Filter sterilize.
Preparation of Medium: Add components, except gelatin solution,
bovine serum albumin solution, and rabbit serum, to distilled/deion-
ized water and bring volume to 1001.0mL. Mix thoroughly. Bring pH
to 7.6 with 5N NaOH. Filter sterilize. Aseptically add 200.0mL of ster-
ile gelatin solution, 143.0mL of sterile bovine serum albumin, and
86.0mL of sterile heat-inactivated rabbit serum. Mix thoroughly. Asep-
tically dispense into sterile tubes or flasks.
Use: For the isolation of Borrelia burgdorferi and other spirochetes.

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