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Gayam, Glydenne Glaire P.

BS Medical Technology 3A
Culture Medium

Bacte MWF 5:30-8:30
August 19, 2015
Use

Preparation

Components

1. Nutrient

Basal Medium

Dissolve 100g agar in 1L DH20. Heat,
dispense, autoclave.

2. Acetate Agar

Differential Medium

3. Alkaline Peptone Water

Enrichment Medium

4. Bismuth Sulfite Agar

Selective Medium

Add components, except sodium
acetate buffer, to distilled/deionized
water and bring volume to 900.0mL.
Mix. Heat and bring to boiling.
Autoclave and cool. Aseptically add
100.0mL of sterile sodium acetate
buffer. Mix.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
thoroughly. Adjust pH to 9.0. Autoclave.
Add components to distilled/deionized
water and bring volume to 1.0L. Mix
and heat until boiling. Boil for 1 min. Do
not autoclave. Cool.

Protein
Sugar
Indicators
Peptone
Sugar
Meat extract
Yeast extract
Sodium acetate buffer

5. Blood Agar, Sheep

Differential Medium

Add components, except sheep blood,
to distilled/deionized water and bring
volume to 950.0mL. Mix. Heat and
bring to boiling. Autoclave and cool
Aseptically add 50.0mL of sterile sheep
blood. Mix. Pour into sterile Petri
dishes.

Peptone
NaCl
Agar
Bismuth sulphite
Casein
Animal tissue
Glucose
Na2HPO4
FeSO4·7H2O
Trypticase soy agar
Brucella agar, or beef heart
infusion with 5% sheep blood

0mL. except L-cysteine solution. Mix.0L. to distilled/deionized water and bring volume to 890. Mix.0mL of L-cysteine solution.0mL of sterile. Heat while stirring and bring to 85°C for 5–10 min. Mix and heat until boiling. Buffered Charcoal-Yeast Extract Agar Enrichment Medium 8.6. Add components to distilled/deionized water and bring volume to 1. except supplement B solution and sheep blood. Pour into sterile Petri dishes or leave in tubes. Mix.0L. Aseptically add 100. Add 4. Cool to 50°C. Add components. Autoclave and cool. Autoclave and pour into Agar Tryptose NaCl Beef Extract Phenylethyl alcohol Yeast extract Agar Charcoal salts Peptone base.0L. defibrinated sheep blood. Mix and pour into sterile Petri dishes with constant agitation to keep charcoal in suspension. enriched with solution of 2% hemoglobin or isovitalex (BBL) NaCl Cornstarch Sheep blood Peptone base (lactose) Eosin Y Methylene Blue indicator Sugar . Add components. to distilled/deionized water and bring volume to 1. Eoisin-Methylene Blue Agar Selective and Differential medium Add components to distilled/deionized water and bring volume to 1. gently heat and bring to boiling. Chocolate Agar Enrichment medium 9. Distribute into tubes or flasks. heat and bring to boil for 1 min. heat and bring to boiling. Aseptically add 10. Autoclave.0mL of sterile supplement B. Distribute into tubes or flasks. Mix and pour into sterile Petridishes or distribute into sterile tubes. Autoclave and cool. Phenylethyl Alcohol Agar Selective Medium 7.

Autoclave. Add components to distilled/deionized water and bring volume to 1. Mix. Pour into sterile Petri dishes or leave in tubes. heat and bring to boiling. Distribute into tubes in 10.0L. MacConkey Agar Selective and Differential medium 15. Kligler’s Iron Agar Differential medium Identify if a gram-negative rod is a glucose or lactose fermenter or both 12. Mix. Mix then heat while stirring until boiling. Lead acetate Agar Peptone Glucose Sugars Indicator Peptone Sulfur source Peptones. yeast extract.0L. Mix. Mix then heat while stirring and bring to boiling. agalactiae 13. Mannitol Salt Agar Selective and Differential medium Add components to distilled/deionized water and bring volume to 1.0L. Allow tubes to cool in a slanted position. Pour into sterile Petri dishes or distribute into sterile tubes. Allow tubes to cool in a slanted position.sterile Petri dishes. Distribute into tubes or flasks.0L. Autoclave. Autoclave. Add components to distilled/deionized water and bring volume to 1. Mix thoroughly.0L.0L. Lysine-Iron Sugar Differential medium Identify species of Enterobacteriaceae 14. Hydrogen Sulfide. Lead Acetate Differential culture 11. Distribute into tubes or flasks. Autoclave and pour into sterile Petri dishes or leave in tubes. Autoclave. dextrose Amino acid Carbohydrate Indicators Sulfur Peptone base Indicator NaCl Bile Salts Peptone base Mannitol Indicator Beef extract . heat and bring to boiling. Lim Broth Enrichment broth Isolate S. Distribute into tubes. Distribute into tubes or flasks. Add components to distilled/deionized water and bring volume to 1. Add components to distilled/deionized water and bring volume to 1. heat while stirring and bring to boiling. 10.0mL volumes. Add components to distilled/deionized water and bring volume to 1.

16.0L. Pour into sterile Petri dishes or leave in tubes. Aseptically combine components. Mix. Add components to distilled/deionized water and bring volume to 1.0L. Pour into sterile Petri dishes or leave in tubes.0-inch butt. Distribute into tubes or flasks. Distribute into tubes or flasks. Add components to distilled/deionized water and bring volume to 1. citrated Yeast dialysate Peptone base broth Sodium biselenite Sugars Indicator Peptone Sulfur source Peptone Casein Sodium chloride . Autoclave and cool in a slanted position to form a 1. Autoclave. Trypticase Soy Agar All-purpose/ Basal medium Add components to distilled/deionized water and bring to 1. Pour into sterile Petri dishes. New York City Medium Selective medium Isolate N. Distribute into tubes or flasks.0mL volumes. Autoclave. heat and bring to boiling.0L. heat and bring to boiling. Triple-Sugar Iron Agar Differential medium Identification of Glucosefermenters from Non-glucosefermenters gram negative rods 20. Selenite Broth Enrichment Broth 19. Mix. Mix. Have all solutions prepared and at 45°– 50°C. Add components to distilled/deionized water and bring volume to 1.0L. Mix. heat and bring to boiling. Mueller-Hinton Agar Transparent medium 17. Do not autoclave. Do not overheat. Distribute into sterile tubes in 10. Pour into sterile Petri dishes or leave in tubes. meningitidis from specimens containing mixed normal flora 18. NaCl Casein Animal Tissue Animal infusion Casein extract Starch Agar Peptone agar base with cornstarch Horse blood cells Horse plasma.Autoclave. Mix. heat and bring to boiling. gonorrhoea and N.

Trypticase Soy Broth All-purpose medium/ Enrichment medium Add components to distilled/deionized water and bring volume to 1. R. Textbook of Diagnostic Microbiology (4th edition). (2014). Soybean Casein NaCl Glucose Dipotassium phosphate . (2010). Handbook of Microbiological Media (4th edition). al (2015). et. Autoclave then mix thoroughly.M. Elsevier Mosby. Tille. Mix. P. Atlas.21. Saunders Elsevier.0L. C. Mahon.M. CRC Press.R. Bailey & Scott’s Diagnostic Microbiology (13th edition). heat and bring to boiling. Distribute into tubes or flasks.