Handbook of Microbiological Media, Fourth Edition part 196 pdf

0.75g Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L.. 0.05g Preparation of Medium: Add components to distilled/deionized water and bring vol

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YI-S Medium 1945

Use: For the cultivation and maintenance of Xanthomonas campestris

and Xanthomonas oryzae.

YGLM (Yeast Glucose Litmus Milk)

Composition per liter:

Glucose 10.0g

Skim milk powder 8.0g

Yeast extract 3.0g

Litmus 0.75g

Preparation of Medium: Add components to distilled/deionized

water and bring volume to 1.0L Mix thoroughly for 15–20 min

Dis-tribute into tubes or flasks Autoclave for 10 min at 10 psi pressure–

115°C Incubate for 1 week at 30°C to check for sterility before use

Use: For the cultivation ofGemella morbillorum.

YGLM with Chalk (Yeast Glucose Litmus Milk with Chalk)

CaCO3 20.0g

Glucose 10.0g

Yeast extract 3.0g

Litmus 0.75g

Skim milk 100.0mL

Preparation of Medium: Add components, except skim milk, to

distilled/deionized water and bring volume to 900.0mL Mix

thorough-ly Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Add

10.0mL of skim milk Distribute into tubes Autoclave for 10 min at 10

psi pressure–115°C Incubate for 1 week at 30°C to check for sterility

before use

Use: For the cultivation ofGemella morbillorum.

YGLPB Medium

Peptone 10.0g

Lab-Lemco 8.0g

Glucose 5.0g

Lactose 5.0g

Yeast extract 3.0g

KH2PO4 2.5g

K2HPO4 2.5g

MgSO4·7H2O 0.2g

MnSO4·4H2O 0.05g

water and bring volume to 1.0L Mix thoroughly Distribute into tubes

or flasks Autoclave for 20 min at 15 psi pressure–121°C

Use: For the cultivation of Lactobacillus delbrueckii.

YGLPB Medium

Compositionper liter:

Peptone 10.0g

Beef extract 8.0g

Glucose 5.0g

Lactose 5.0g

Yeast extract 3.0g

K2HPO4 2.5g

KH2PO4 2.5g

MgSO4·7H2O 0.2g MnSO4·4H2O 0.05g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.8 Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure– 121°C

Use: For the cultivation of Carnobacterium divergens, Carnobacterium piscicola, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactococ-cus lactis, StreptococLactococ-cus ferus, and StreptococLactococ-cus sobrinus.

YGLPB Medium

Peptone 1.0g Lab Lemco (meat extract) 0.8g Glucose 0.5g Lactose 0.5g Yeast extract 0.3g

K2HPO4 0.25g

KH2PO4 0.25g MgSO4·7H2O 0.02g MnSO4·4H2O 5.0mg

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Distribute into tubes

Use: For the cultivation of Carnobacterium gallinarum, Carnobacterium mobile, Enterococcus dispar, Lactobacillus fructivorans, Leuconostoc carnosum, Leuconostoc gelidum, and Vagococcus salmoninarum.

YGLPB Medium without Lactose

Peptone 10.0g Lab Lemco 8.0g Yeast extract 3.0g Glucose 5.0g

KH2PO4 2.5g

K2HPO4 2.5g MgSO4·7H2O 0.2g MnSO4·4H2O 0.05g

Use: For the cultivation of Vagococcus fluvialis and Vagococcus sal-moninarum

YI-S Medium

YI broth 880.0mL Bovine serum, heat inactivated 100.0mL Vitamin mixture 18 20.0mL

Source: Vitamin mixture 18 is available from Bio-fluids, Inc., Rock-ville, MD

YI Broth:

YI base stock 780.0mL 10X Glucose buffer stock 100.0mL

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1946 YM-1L Broth

YI Base Stock:

Compositionper 780.0mL:

Yeast extract 30.0g

L-Cysteine·HCl 1.0g

NaCl 1.0g

Ascorbic acid 0.2g

Ferric ammonium citrate 228.0mg

10X Glucose Buffer Stock:

Composition per 100.0.0mL:

Glucose 10.0g

K2HPO4 1.0g

KH2PO4 0.6g

Preparation of 10X Glucose Buffer Stock: Add components to

distilled/deionized water and bring volume to 100.0mL Mix

thorough-ly Filter sterilize

Preparation of YI Base Stock: Add components to 600.0mL of

distilled/deionized water Mix thoroughly Bring volume to 780.0mL

with distilled/deionized water Adjust pH to 6.8 with 1N NaOH

Dis-tribute in 78.0mL aliquots to 100.0mL screw-capped bottles

Auto-clave for 15 min at 15 psi pressure–121°C Cool to room temperature

Preparation of YI Broth: Aseptically add 10.0mL of 10X glucose

buffer stock to 78.0mL of cooled YI base stock Adjust osmolarity with

NaCl to 380.0milliosmols/kg

Preparation of Medium: Aseptically add 2.0mL of vitamin

mix-ture 18 and 10.0mL of heat-inactivated bovine serum to 88.0mL of YI

broth Distribute in 13.0mL aliquots to 16 x 125mm screw-capped test

tubes Store at 4°C in the dark with the caps screwed on tightly Use

within 96 hr

Use: For the cultivation of Entamoeba species.

YM Agar

See: Yeast Malt Extract Agar

YM Broth

See: Yeast Malt Extract Broth

YM Broth with 0.5%CaCO 3

YM Broth with 2.0%CaCO 3

YM Broth with Glucose

See: Yeast Malt Extract Broth with Glucose

YM Broth with 1.0% Methanol

See: Yeast Malt Extract Broth with 1.0% Methanol

YM Broth with 18% NaCl

See: Yeast Malt Extract Broth with 18% NaCl

YM Broth with 40% Sucrose

See: Yeast Malt Extract Broth with 40% Sucrose

YM Broth with 70% Sucrose

See: Yeast Malt Extract Broth with 70% Sucrose

YM Catalase Agar

See: Yeast Malt Extract Catalase Agar

YM-1L Broth

Sodium lactate 30.0g Mycological peptone 10.0g Succinic acid 10.0g NaOH 6.0g Yeast extract 5.0g Adenine 0.01g Uracil 0.01g Yeast nitrogen base solution 100.0mL

Yeast Nitrogen Base Solution:

(NH4)2SO4 5.0g

KH2PO4 1.0g MgSO4·7H2O 0.5g NaCl 0.1g CaCl2·2H2O 0.1g DL-Methionine 0.02g DL-Tryptophan 0.02g L-Histidine·HCl 0.01g Inositol 2.0mg

KI 0.1mg

H3BO3 0.5mg ZnSO4·7H2O 0.4mg MnSO4·4H2O 0.4mg Thiamine·HCl 0.4mg Pyroxidine·HCl 0.4mg Niacin 0.4mg Calcium pantothenate 0.4mg

p-Aminobenzoic acid 0.2mg

Riboflavin 0.2mg FeCl3 0.2mg

Na2MoO4·4H2O 0.2mg CuSO4·5H2O 0.04mg Folic acid 2.0μg Biotin 2.0μg

Preparation of Yeast Nitrogen Base Solution: Add components

to distilled/deionized water and bring volume to 1.0L Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except yeast nitrogen base solution, to distilled/deionized water and bring volume to 900.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure– 121°C Aseptically add 1.0L of sterile yeast nitrogen base solution Mix thoroughly Aseptically distribute into sterile tubes or flasks

Use: For the cultivation ofSaccharomyces cerevisiae.

YM Medium

See: Universal Agar for Yeasts

YM Medium (DSMZ Medium 1070)

Mannitol 10.0g Yeast extract 0.5g

K2HPO4 0.5g NaCl 0.2g CaCl2·2H2O 0.2g MgSO4·7H2O 0.1g

pH 7.0 ± 0.2 at 25°C

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YMF Broth 1947

water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.0

Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–

121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of Labrys miyagiensis.

YM5 with 10% Sorbitol

Sorbitol 100.0g

Glucose 10.0g

Succinic acid 10.0g

NaOH 6.0g

Peptone 2.0g

Yeast extract 1.0g

Adenine 0.01g

Uracil 0.01g

Yeast nitrogen base solution 100.0mL

pH 5.8 ± 0.2 at 25°C

Yeast Nitrogen Base Solution:

(NH4)2SO4 5.0g

KH2PO4 1.0g

MgSO4·7H2O 0.5g

NaCl 0.1g

CaCl2·2H2O 0.1g

DL-Methionine 0.02g

DL-Tryptophan 0.02g

L-Histidine·HCl 0.01g

Inositol 2.0mg

KI 0.1mg

H3BO3 0.5mg

ZnSO4·7H2O 0.4mg

MnSO4·4H2O 0.4mg

Thiamine·HCl 0.4mg

Pyroxidine·HCl 0.4mg

Niacin 0.4mg

Calcium pantothenate 0.4mg

Riboflavin 0.2mg

FeCl3 0.2mg

Na2MoO4·4H2O 0.2mg

CuSO4·5H2O 0.04mg

Folic acid 2.0μg

Biotin 2.0μg

Source: Yeast nitrogen base is available as a premixed powder from

BD Diagnostic Systems

Preparation of Yeast Nitrogen Base Solution: Add components

to distilled/deionized water and bring volume to 100.0mL Mix

thor-oughly Filter sterilize

Preparation of Medium: Add components, except yeast nitrogen

base solution, to distilled/deionized water and bring volume to

900.0mL Mix thoroughly Autoclave for 15 min at 15 psi pressure–

121°C Cool to room temperature Aseptically add 100.0mL of sterile

yeast nitrogen base solution to the cooled sterile basal medium Mix

thoroughly Adjust final pH to 5.8 Distribute into sterile flasks or

tubes

Use: For the cultivation of a sorbitol-utilizing fungus

YMA Agar

Agar 15.0g Mannitol 10.0g CaCO3 4.0g

KH2PO4 0.5g Yeast extract 0.4g MgSO4·7H2O 0.2g NaCl 0.1g

pH 6.8 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Gently heat and bring

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of Rhizobium fredii, Rhizobium galegae, Rhizo-bium huakuii, RhizoRhizo-bium leguminosarum, RhizoRhizo-bium loti, RhizoRhizo-bium meliloti, Rhizobium phaseoli, and Rhizobium trifolii.

YMA Medium (DSMZ Medium 1031)

Agar 20.0g Mannitol 10.0g Yeast extract 0.3g MgSO4·7H2O 0.2g

K2HPO4 0.2g NaCl 0.05g

pH 7.0 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 7.0 Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure– 121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of Phyllobacterium trifolii

YMF Agar

Agar 20.0g Peptone 5.0g Sugar, brown 3.0g Malt extract 3.0g Yeast extract 3.0g

pH 6.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.2 Gently heat and bring to boiling Distribute into tubes or flasks Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

Use: For the cultivation of various fungi

YMF Broth

Peptone 5.0g Sugar, brown 3.0g Malt extract 3.0g Yeast extract 3.0g

pH 6.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.2

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1948 YNA Medium

121°C

Use: For the cultivation of various fungi

YNA Medium (Yeast Extract Nutrient Agar Medium)

Agar 15.0g

NaCl 5.0g

Peptone 5.0g

Meat extract 4.0g

Yeast extract 2.5g

pH 7.0 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Gently heat and bring

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15

psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the isolation and cultivation of Kurthia species according to

the agar streak method

YNG Medium (Yeast Extract Nutrient Gelatin Medium)

Gelatin 100.0g

NaCl 5.0g

Peptone 5.0g

Meat extract 4.0g

Yeast extract 2.5g

pH 7.0 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Gently heat until

dis-solved Distribute into tubes or flasks Autoclave for 30 min at 10 psi

pressure–115°C

Use: For the isolation and cultivation of Kurthia species using the

gel-atin streak method

Yolk Milk Medium (YOM)

Egg yolk 500.0mL

Milk 500.0mL

Egg Yolk:

Composition per 500.0mL:

Chicken egg yolks variable

Preparation of Egg Yolk: Soak eggs with 1:100 dilution of

saturat-ed mercuric chloride solution for 1 min Crack eggs and separate yolks

from whites Add sufficient egg yolk to bring volume to 500.0mL Mix

thoroughly

Preparation of Milk: Autoclave 500.0mL of milk for 20 min at 15

psi pressure–115°C

Preparation of Medium: Combine 500.0mL of sterile egg yolk and

500.0mL of sterile milk Mix thoroughly Distribute into sterile tubes

or flasks Heat to 95°C for 20–25 min

Use: For the cultivation of Condiobolus obscurus.

Yopp’s Medium

NaCl 116.88g MgCl2·6H2O 10.68g MgSO4·7H2O 10.0g KCl 2.0g CaNO3·4H2O 1.0g Glycyl-glycine buffer 0.5g

K2HPO4·3H2O 0.065g Ferric EDTA 5.0mg Trace metals solution 1.0mL

pH 7.8 ± 0.2 at 25°C

Trace Metals Solution:

MnCl2·4H2O 2.0g

H3BO3 0.5g ZnNO3·6H2O 0.5g Co(NO3)2·6H2O 0.025g CuCl2·2H2O 0.025g

Na2MoO4·2H2O 0.025g VOSO4·6H2O 0.025g HCl 3.0mL

Preparation of Trace Metals Solution : Add components to

dis-tilled/deionized water and bring volume to 1.0L Mix thoroughly

to boiling Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C

Use: For the isolation and cultivation of halophilic cyanobacteria

YP87 Medium

Na2SO4 4.0g NaHCO3 1.3g KCl 0.5g Yeast extract 0.5g MgCl2·6H2O 0.4g

NH4Cl 0.25g

L-Ascorbic acid 0.2g

Na2HPO4 0.2g Sodium thioglycolate 0.2g CaCl2·2H2O 0.15g Resazurin 1.0mg Modified Wolfe’s mineral solution 10.0mL Wolfe’s vitamin solution 10.0mL Sodium lactate, 60% syrup 3.0mL

pH 7.5 ± 0.2 at 25°C

Modified Wolfe’s Mineral Solution:

MgSO4·7H2O 3.0g Nitrilotriacetic acid 1.5g NaCl 1.0g MnSO4·H2O 0.5g CaCl2 0.1g CoCl2·6H2O 0.1g FeSO4·7H2O 0.1g ZnSO4·7H2O 0.1g AlK(SO4)2·12H2O 0.01g CuSO4·5H2O 0.01g

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YPDP Medium with 5´-TMP 1949

H3BO3 0.01g

Na2MoO4·2H2O 0.01g

Na2SeO3 0.01g

NaWO4·2H2O 0.01g

NiC12·6H2O 0.01g

Preparation of Modified Wolfe’s Mineral Solution: Add

nitrilotriacetic acid to 500.0mL of distilled/deionized water Adjust pH

to 6.5 with KOH Add remaining components one at a time Add

dis-tilled/deionized water to 1.0L Adjust pH to 6.8

Wolfe’s Vitamin Solution:

Pyridoxine·HCl 10.0mg

Lipoic acid 5.0mg

Nicotinic acid 5.0mg

Riboflavin 5.0mg

Thiamine·HCl 5.0mg

Calcium DL-pantothenate 5.0mg

Biotin 2.0mg

Folic acid 2.0mg

Vitamin B12 0.1mg

Preparation of Wolfe’s Vitamin Solution: Add components to

distilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Prepare and dispense medium under 100%

N2 Add components, except L-ascorbic acid, NaHCO3, and sodium

thio-glycolate, to distilled/deionized water and bring volume to 1.0L Mix

thoroughly Adjust pH to 7.5 Gently heat and bring to boiling Cool

while sparging with 100% N2 Add L-ascorbic acid, NaHCO3, and

sodi-um thioglycolate Mix thoroughly Sparge with 100% N2 Distribute into

tubes or bottles Autoclave for 15 min at 15 psi pressure–121°C

Use: For the cultivation of Thermodesulfovibrio yellowstonii

YPAD Medium for MAK Mutants of Saccharomyces

Peptone 20.0g

Glucose 20.0g

Agar 20.0g

Yeast extract 10.0g

Adenine sulfate 0.4g

Use: For the cultivation and maintenance of a variety of yeasts, including

Candida albicans, Candida boidinii, Candida pintolopesii,

Saccharomy-ces cerevisiae, and SchizosaccharomySaccharomy-ces pombe

YPC Medium

Agar 15.0g

Proteose peptone 15.0g

Yeast extract 5.0g

KH2PO4 4.0g

Sucrose 2.5g

Glucose 2.0g

L-Cystine 0.5g

Na2SO3 0.2g

pH 7.2 ± 0.2 at 25°C

Use: For the cultivation of Pasteurella multocida.

YPD Medium (DSMZ Medium 393)

Peptone 20.0g Glucose 20.0g Yeast extract 10.0g

pH 6.5 ± 0.2 at 25°C

Use: For the isolation and cultivation of Yarrowia lipolytica (Candida lipolytica), Kluyveromyces spp., Saccharomyces spp., Pichia spp., and Candida spp.

YPDA (Yeast Peptone Dextrose Agar)

Agar 20.0g Glucose 20.0g Peptone 20.0g Yeast extract 10.0g Adenine sulfate 0.4g

Use: For the cultivation of Taphrina populina

YPDP Medium with 5´-TMP

Glucose 20.0g Peptone 20.0g Agar 15.0g Yeast extract 10.0g

KH2PO4 1.5g Thymidine-5´-monophosphate solution 10.0mL

Thymidine-5´-Monophosphate Solution:

Thymidine-5´-monophosphate 100.0mg

Preparation of Thymidine-5´-Monophosphate Solution: Add thymidine-5´-monophosphate to distilled/deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except thymidine-5´-monophosphate solution, to distilled/deionized water and bring vol-ume to 990.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C Asepti-cally add 10.0mL of sterile thymidine-5´-monophosphate solution Mix thoroughly Pour into sterile Petri dishes or aseptically distribute into sterile tubes

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1950 YPG Agar with 2% Sodium Chloride

Use: For the cultivation and maintenance of Saccharomyces

cerevi-siae.

YPG Agar with 2% Sodium Chloride

Agar 20.0g

Glucose 20.0g

NaCl 20.0g

Peptone 10.0g

Yeast extract 10.0g

pH 5.2 ± 0.2 at 25°C

Gently heat and bring to boiling Distribute into tubes or flasks

Auto-clave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes

or leave in tubes

YPG Medium (DSMZ Medium 1017)

Glucose 70.0gl

Yeast extract 10.0g

Peptone 10.0g

pH 6.0 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Adjust pH to 6.0 with

dilute HCl Distribute into tubes or flasks Autoclave for 15 min at 15

psi pressure–121°C

Use: For the cultivation of Saccharibacter floricola.

YPG Medium (DSMZ Medium 1172)

Yeast extract 1.0g

Peptone 1.0g

Glucose 1.0g

pH 5.7 ± 0.2 at 25°C

water and bring volume to 1.0L Mix thoroughly Adjust pH to 5.6–6.0

with dilute HCl Distribute into tubes or flasks Autoclave for 15 min

at 15 psi pressure–121°C

Use: For the cultivation of Asticcacaulis benevestitus

YPGA (DSMZ Medium 1015)

Agar 15.0g

Yeast extract 7.0g

Peptone 7.0g

Glucose 7.0g

pH 7.3 ± 0.2 at 25°C

Use: For the cultivation of Stenotrophomonas maltophilia.

Use: For the cultivation of a variety of yeasts and other fungi

YPM Agar

Mannitol 25.0g Agar 12.0g Yeast extract 5.0g Peptone 3.0g

to boiling Do not adjust pH Distribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–121°C Pour into sterile Petri dishes or leave in tubes

Use: For the cultivation of Acetobacter aceti, Acetobacter pasteurianus, Acetobacter xylinum, Frateuria aurantia, and Pseudomonas aeruginosa.

YPNC Medium

Agar 18.0g NaCl 2.92g

KH2PO4 0.596g Yeast extract 0.5g Sodium hydrogen glutamate (pH 6.0) 0.37g

K2HPO4 0.107g

NH4Cl 0.107g MgSO4·7H2O 0.049g Glucose solution 10.0mL Trace metals solution 1.0mL

pH 6.0 ± 0.2 at 25°C

Glucose Solution:

Glucose 4.0g

Preparation of Glucose Solution: Add glucose to distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Filter steril-ize

Trace Metals Solution:

EDTA 50.0g ZnSO4·7H2O 22.0g CaCl2 5.54g MnCl2·4H2O 5.06g FeSO4·7H2O 4.99g (NH4)6Mo7O14·H2O 1.10g CoSO4·5H2O 1.57g CoCl2·6H2O 1.61g

Preparation of Trace Metals Solution: Add components, one at

a time, to distilled/deionized water and bring volume to 1.0L Mix thor-oughly Filter sterilize

Preparation of Medium: Add components, except glucose solu-tion and trace metals solusolu-tion, to distilled/deionized water and bring volume to 989.0mL Mix thoroughly Gently heat and bring to boiling Adjust pH to 6.0 with KOH Autoclave for 15 min at 15 psi pressure– 121°C Cool to 50°C Aseptically add 10.0mL of sterile glucose solu-tion and 1.0mL of sterile trace metals solusolu-tion Mix thoroughly Pour into sterile Petri dishes or aseptically distribute into sterile tubes

Use: For the cultivation and maintenance of a variety of Cryptococcus

species.

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YPSC Medium 1951

YPS Medium (DSMZ Medium 990)

Sea salts, Sigma 35.0g

Sulfur, elemental 5.0g

PIPES (piperazine-N,N'-bis[2-ethane-sulfonic acid]) 3.46g

NH4Cl 0.5g

KH2PO4 0.35g

CaCl2·2H2O 0.2g

FeCl3·6H2O 6.7mg

Na2WO4 2.9mg

Resazurin 0.1mg

Yeast extract solution 10.0mL

Peptone solution 10.0mL

Sulfide solution 5.0mL

pH 6.8 ± 0.2 at 25°C

Sulfide Solution :

Na2S·9H2O 0.5g

Preparation of Sulfide Solution: Add Na2S·9H2O to

distilled/de-ionized water and bring volume to 10.0mL Mix thoroughly Autoclave

under 100% N2 for 15 min at 15 psi pressure–121°C Cool to room

temperature Adjust pH to 7.0

Yeast Extract Solution:

Yeast extract 1.0g

Preparation of Yeast Extract Solution: Add yeast extract to

dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly

Autoclave for 15 min at 15 psi pressure–121°C Cool to room

temper-ature Sparge with 100% N2

Peptone Solution:

Peptone 4.0g

Preparation of Peptone Solution: Add peptone to

distilled/deion-ized water and bring volume to 10.0mL Mix thoroughly Autoclave for

15 min at 15 psi pressure–121°C Cool to room temperature Sparge

with 100% N2

Preparation of Medium: Add components, except yeast extract,

peptone, and sulfide solutions, to distilled/deionized water and bring

volume to 975.0mL Mix thoroughly Gently heat and bring to boiling

Cool to room temperature while sparging with 100% N2 Adjust pH to

6.8 Prepare the medium without the yeast extract, peptone, and sodium

sulfide Boil the medium and cool under nitrogen Adjust the pH to 6.8

Dispense into Hungate tubes or serum bottles under a nitrogen

atmo-sphere Sterilize the medium at 100°C for 3 hr on 3 consecutive days

Aseptically add the peptone and yeast extract solutions Adding the

sterile, neutralized sulfide solution to an end concentration of 0.025%

Final pH should be 6.8

Use: For the cultivation of Thermococcus marinus.

YPS Medium (DSMZ Medium 1168)

Sea salts, Sigma 25.0g

Yeast extract 4.0g

Peptone 2.0g

pH 7.5 ± 0.2 at 25°C

Use: For the cultivation of Owenweeksia hongkongensis.

YPSC Agar (Yeast Extract Peptone Sulfate Cysteine Agar)

Agar 15.0g Yeast extract 1.0g Peptone 1.0g Sodium acetate·3H2O 0.5g MgSO4·7H2O 0.25g CaCl2·2H2O 0.25g L-Cysteine·HCl·H2O 0.05g

pH 7.5 ± 0.2 at 25°C

psi pressure–121°C Adjust pH to 7.5 with sterile 10M NaOH Pour

into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Bdellovibrio species.

YPSC Agar, Cation-Supplemented

Sodium acetate·3H2O 50.0g Agar 15.0g Peptone 10.0g Yeast extract 10.0g MgSO4·7H2O 0.74g CaCl2·2H2O 0.29g L-Cysteine·HCl·H2O 0.05g Bacitracin solution 10.0mL

Bacitracin Solution:

Bacitracin 6,000U

Preparation of Bacitracin Solution: Add bacitracin to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except bacitracin solu-tion, to distilled/deionized water and bring volume to 990.0mL Mix thoroughly Gently heat and bring to boiling Autoclave for 15 min at

15 psi pressure–121°C Cool to 45°–50°C Aseptically add sterile bac-itracin solution Mix thoroughly Pour into sterile Petri dishes or dis-tribute into sterile tubes

Use: For the cultivation and enumeration of Bdellovibrio species.

YPSC Medium (Yeast Extract Peptone Sulfate Cysteine Medium)

Yeast extract 1.0g Peptone 1.0g Sodium acetate·3H2O 0.5g MgSO4·7H2O 0.25g CaCl2·2H2O 0.25g L-Cysteine·HCl·H2O 0.05g

pH 7.5 ± 0.2 at 25°C

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1952 YPSC Soft Agar

or flasks Autoclave for 15 min at 15 psi pressure–121°C Adjust pH to

7.5 with sterile 10M NaOH

Use: For the cultivation and enumeration of Bdellovibrio species.

YPSC Soft Agar (Yeast Extract Peptone Sulfate Cysteine Soft Agar)

Agar 6.0g

Yeast extract 1.0g

Peptone 1.0g

Sodium acetate·3H2O 0.5g

MgSO4·7H2O 0.25g

CaCl2·2H2O 0.25g

L-Cysteine·HCl·H2O 0.05g

pH 7.5 ± 0.2 at 25°C

psi pressure–121°C Adjust pH to 7.5 with sterile 10M NaOH Pour

into sterile Petri dishes or leave in tubes

Use: For the cultivation and maintenance of Bdellovibrio species.

YpSs Agar

Agar 15.0g

Soluble starch 15.0g

Yeast extract 4.0g

K2HPO4 1.0g

MgSO4·7H2O 0.5g

Use: For the cultivation of bacteria that can utilize starch as a carbon

source

YPSS, Emerson Agar

Agar 15.0g

Soluble starch 15.0g

Yeast extract 4.0g

K2HPO4 1.0g

MgSO4·7H2O 0.5g

Use: For the cultivation and maintenance of Allomyces javanicus,

Melanospora tiffanii, and Sporothrix schenckii.

YSP Agar

Sucrose 20.0g

Agar 12.0g

Peptone 10.0g Yeast extract 5.0g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation of Xanthomonas albilineans.

YT HiVeg Broth

Plant hydrolysate 16.0g Yeast extract 10.0g NaCl 5.0g

pH 7.2 ± 0.2 at 25°C

Source: This medium is available as a premixed powder from Hi-Media

Use: For the cultivation of Escherichia coli.

YT Medium (Yeast Extract Tryptone Medium)

Pancreatic digest of casein 8.0g Yeast extract 5.0g NaCl 5.0g

pH 7.2 ± 0.2 at 25°C

Use: For the cultivation of Escherichia coli.

YTG Medium

Tryptone 10.0g

Na2CO3 5.3g Yeast extract 5.0g

Na2HPO4·2H2O 0.356g

L-Cysteine·HCl 0.2g

Na2S·9H2O 0.2g KCl 0.075g Resazurin 1.0mg Glucose solution 20.0mL

pH 10.1 ± 0.2 at 25°C

Glucose Solution:

D-Glucose 3.0g

Preparation of Glucose Solution: Add glucose to distilled/deion-ized water and bring volume to 20.0mL Mix thoroughly Sparge with 100% N2 Autoclave for 15 min at 15 psi pressure–121°C

Preparation of Medium: Prepare and dispense medium under 100% N2 Add components, except glucose solution, to distilled/deion-ized water and bring volume to 980.0L Mix thoroughly Sparge with 100% N2 for 30 min Anaerobically distribute 9.8mL volumes into tubes Autoclave for 15 min at 15 psi pressure–121°C Aseptically and

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Zavarzinella formosa Medium 1953

anaerobically add 0.2mL of sterile glucose solution to each tube

Ad-just pH to 10.1 with sterile anaerobic 3N NaOH solution.

Use: For the cultivation of Clostridium paradoxum and Clostridium

thermoalcaliphilum.

YTN Medium (Yeast Extract Tryptone NaCl Medium)

NaCl 30.0g

Agar 15.0g

Yeast extract 10.0g

Pancreatic digest of casein 10.0g

Glucose 1.0g

Trace elements solution 1.0mL

pH 7.2 ± 0.2 at 25°C

Trace Elements Solution:

H3BO3 2.85g

MnCl2·4H2O 1.8g

Sodium tartrate 1.77g

FeSO4 1.36g

CoCl2·6H2O 0.04g

CuCl2·2H2O 0.027g

Na2MoO4·2H2O 0.025g

ZnCl2 0.021g

pH 7.2 ± 0.2 at 25°C

Preparation of Trace Elements Solution: Add components to

distilled/deionized water and bring volume to 1.0L Mix thoroughly

Preparation of Medium: Add components to tap water and bring

volume to 1.0L Mix thoroughly Gently heat and bring to boiling

Dis-tribute into tubes or flasks Autoclave for 15 min at 15 psi pressure–

Use: For the cultivation of ATCC strain 21588

YTSS Medium, Half Strength

(DSMZ Medium 974)

Sea salts 20.0g

Yeast extract 2.0g

Tryptone 1.25g

pH 7.0 ± 0.2 at 25°C

Use: For the cultivation of Roseovarius nubinhibens and Silicibacter

pomeroyi.

Z Agar

Agar 16.0

K2HPO4 5.0g

K2SO4 2.0g

KH2PO4 1.0g

MgSO4·7H2O 0.05g

Acetamide solution 40.0mL

pH 7.2 ± 0.2 at 25°C

Acetamide Solution:

Acetamide 10.0g

Preparation of Acetamide Solution: Add acetamide to distilled/ deionized water and bring volume to 100.0mL Mix thoroughly Filter sterilize

Preparation of Medium: Add components, except acetamide solu-tion, to distilled/deionized water and bring volume to 960.0mL Mix thoroughly Autoclave for 20 min at 15 psi pressure–121°C Cool to 70°C Aseptically add 40.0mL sterile acetamide solution Mix thor-oughly Pour into sterile Petri dishes Dry plates at 37°C for 30 min

Use: For the isolation of Pseudomonas aeruginosa from milk.

Z Broth

Acetamide 5.0g

K2HPO4 5.0g

KH2PO4 3.0g KNO3 1.0g

K2S4O6 1.0g MgSO4·7H2O 0.05g

pH 7.2 ± 0.2 at 25°C

Preparation of Medium: Add components to distilled/deionized wa-ter and bring volume to 1.0L Mix thoroughly Gently heat and bring to boiling Cool Aseptically distribute 10.0mL volumes into test tubes con-taining inverted Durham tubes Heat for 15 min at 0 psi pressure–100°C

Use: For the cultivation of Pseudomonas aeruginosa from milk.

Z Medium

Casein hydrolysate 10.0g NaCl 10.0g Yeast extract 5.0g Glucose 1.0g CaCl2·2H2O 0.367g

Use: For the cultivation of Alcaligenes eutrophus.

Zavarzinella formosa Medium

(DSMZ Medium 1196)

N-acetylglucosamine 1.0g Glucose 0.5g

KH2PO4 0.1g Peptone 0.1g Yeast extract 0.1g MgSO4·7H2O 0.1g Casamino acids 0.1g CaCl2·2H2O 0.05g NaCl 0.01g

“Metals 44” 1.0mL

pH 5.9 ± 0.2 at 25°C

“Metals 44”:

ZnSO4·7H2O 1.095g FeSO4·7H2O 0.5g

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1954 ZF2 Medium

Sodium EDTA 0.25g

MnSO4·H2O 0.154g

CuSO4·5H2O 39.2mg

Co(NO3)2·6H2O 24.8mg

Na2B4O7·10H2O 17.7mg

Preparation of “Metals 44”: Add sodium EDTA to

distilled/de-ionized water and bring volume to 90.0mL Mix thoroughly Add a few

drops of concentrated H2SO4 to retard precipitation of heavy metal

ions Add remaining components Mix thoroughly Bring volume to

100.0mL with distilled/deionized water

water and bring volume to 1.0L Mix thoroughly Adjust pH to 5.8–6.0

121°C

Use: For the cultivation of Zavarzinella formosa.

ZF2 Medium (DSMZ Medium 943)

NaHCO3 3.8g

Yeast extract 3.0g

(NH4)HCO3 0.45g

MgSO4·6H2O 0.13g

CaCl2·2H2O 0.12g

Resazurin 0.5mg

Phosphate buffer 10.0mL

Glycine solution 5.0mL

Arginine solution 5.0mL

Na2S·9H2O solution 5.0mL

Dithionite solution 1.0mL

Seven vitamin solution 1.0mL

Trace elements solution SL-10 1.0mL

Selenite-tungstate solution 1.0mL

pH 7.3 ± 0.2 at 25°C

Arginine Solution:

Arginine-HCl 3.5g

Preparation of Arginine Solution: Add arginine-HCl to distilled/

deionized water and bring volume to 10.0mL Mix thoroughly Filter

sterilize

Glycine Solution:

Glycine 15.0g

Preparation of Glycine Solution: Add glycine to

distilled/deion-ized water and bring volume to 100.0mL Mix thoroughly Sparge with

100% N2 Autoclave for 15 min at 15 psi pressure–121°C Cool to

room temperature

Na 2 S·9H 2 O Solution :

Na2S·9H2O 0.3g

Preparation of Na 2 S·9H 2 O Solution: Add Na2S·9H2O to

dis-tilled/deionized water and bring volume to 10.0mL Mix thoroughly

Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool

to room temperature

Dithionite Solution :

Na2-dithionite 0.25g

Preparation of Dithionite Solution : Add Na2-dithionite to distilled/ deionized water and bring volume to 10.0mL Mix thoroughly Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C Cool to 25°C

Seven Vitamin Solution:

Pyridoxine hydrochloride 300.0mg Thiamine-HCl·2H2O 200.0mg Nicotinic acid 200.0mg Vitamin B12 100.0mg Calcium pantothenate 100.0mg

D(+)-Biotin 20.0mg

Preparation of Seven Vitamin Solution: Add components to dis-tilled/deionized water and bring volume to 1.0L Sparge with 100% N2 Mix thoroughly Filter sterilize

Phosphate Buffer:

Na2HPO4·12H2O 43.0g

KH2PO4 5.44g

Preparation of Phosphate Buffer: Add components to distilled/ deionized water and bring volume to 1.0L Mix thoroughly Adjust pH

to 7.3 Autoclave for 15 min at 15 psi pressure–121°C Cool to 50°C

Trace Elements Solution SL-10:

FeCl2·4H2O 1.5g CoCl2·6H2O 190.0mg MnCl2·4H2O 100.0mg ZnCl2 70.0mg

Na2MoO4·2H2O 36.0mg NiCl2·6H2O 24.0mg

H3BO3 6.0mg CuCl2·2H2O 2.0mg HCl (25% solution) 10.0mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O

to 10.0mL of HCl solution Mix thoroughly Add distilled/deionized water and bring volume to 1.0L Add remaining components Mix thor-oughly Sparge with 80% N2 + 20% CO2 Autoclave for 15 min at 15 psi pressure–121°C

Selenite-Tungstate Solution Compositionper liter:

NaOH 0.5g

Na2WO4·2H2O 4.0mg

Na2SeO3·5H2O 3.0mg

Preparation of Selenite-Tungstate Solution: Add components

to distilled/deionized water and bring volume to 1.0L Mix thoroughly Sparge with 100% N2 Filter sterilize

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas mixture Add components, except phosphate buffer, glycine solution, arginine solution, dithionite solution, seven vitamin so-lution, and Na2S·9H2O solution, to distilled/deionized water and bring volume to 973.0mL Mix thoroughly Equilibrate with 80% N2 + 20%

CO2 to reach pH 7.3 Autoclave for 15 min at 15 psi pressure–121°C Cool to 25°C Aseptically and anaerobically add 10.0mL phosphate buf-fer, 5.0mL glycine solution, 5.0mL arginine solution, 1.0mL dithionite solution, 1.0mL seven vitamin solution, and 5.0mL Na2S·9H2O solution Mix thoroughly Aseptically and anaerobically distribute into sterile tubes or flasks

Use: For the cultivation of Sedimentibacter saalensis.

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