Prosedur PembuatanReagent Uji Biokimia

Untuk Analisa Mikrobiologi .

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For detection of acetylmethylcarbinolSolution A Alpha-naphthol = 5,0 g Ethanol, Absolute = 95,0 gNote : Dissolve the alpha-naphthol in the

ethanol with constant stirring.Solution B Potassium Hydroxide = 40,0 g Creatine = 0,3 g Distilled Water = 100,0 g

Note : Dissolve the potassium hydroxide in 75 ml of destilled water. The solution will become warm. Allow to cool to room temperature. Add the creatine and stir to dissolve. Add the remaining water. Store in a refrigerator.

For ames testl- Histidine HCl = 0,5 mmBiotin = 0,5 mmDistilled Water = 10,0 ml

For immunofluorescence Glycerin = 90,0 ml Phosphate Buffered Saline = 10,0 ml

For detection of nitrates.Dissolve 0,7 g diphenylamine in a mixture of

60 ml concentrate sulfuric acid and 28,8 ml of distilled water. Cool and slowly add 11,3 ml of concentrated hydrochloric acid. Allow to stand for a 12 hours. Sedimentation indicates that the reagent is saturated.

Ferric Chloride = 10,0 g Destilled water = 100,0 ml

Iodine = 1,0 g Potassium iodide = 2,0 g Distilled water = 300,0 ml

For detection of catalase activity.Note : Refrigerate when not in use.

For detectin of indole.p-Dimethylaminobenzaldehyde = 5,0 gAmly alcohol = 75,0 mlHydrochloric Acid (concentrated) = 25,0 mlNote : Dissolve the p-Dimethylaminobenzaldehyde in the amly alcohol. Add the hydrochloric Acid.

For API ® Staph – ident procedure. Prepare 1% aquerus barium chloride and 1% aqueous sulfuric acid solutions. Using the following table, add the amounts of barium chloride and sulfuric acid to clean 15- X 150-mm screw-capped test tubes. Label the tubes 1 through 10

Tube Barium Chloride 1%

(ml)

Sulfuric Acid 1%

(ml)

Corresponding Approximate Density of

Bacteria (million/ml)

1 0,1 9,9 300

2 0,2 9,8 600

3 0,3 9,7 900

4 0,4 9,6 1,200

5 0,5 9,5 1,500

6 0,6 9,4 1,800

7 0,7 9,3 2,100

8 0,8 9,2 2,400

9 0,9 9,1 2,700

10 1,0 9,0 3,000

For microscopic observation of protozoa Methl cellulose = 10,0 gDistilled water 90,0 ml

For detection of acid Methyl red = 0,1 g Ethyl alcohol 300,0 ml Distilled water = 200,0 mlNote : Dissolve the methyl red in the 95

% ethyl alcohol. Dilute to 500 ml with ditilled water

For detection of ammonia Potassium iodine = 50,0 g Distilled water (ammonia-free) = 35,0 mlAdd saturated aqueous solution of mercuric

chloride until a slight precipitate persists. Potassium hydroxide (50 % aqueous) =

400,0 mlNote : Dilute to 1000 ml with ammonia-free

distilled water. Let stand for 1 week, decant supernatant liquid, and store in a tightly capped amber bottle.

For detection of nitrites. Solution A, Sulfanilic acid :

Sulfanilic acid = 8,0 g Acetic acid, 5 N: 1 part glacial acetic acid

to 2,5 parts Distilled water = 1000,0 mlSolutiob B, alpha-naphthylamine Alpha-naphthylamine = 5,0 g Acetic acid, 5 N = 1000,0 ml

For enzyme induction 0,1 M sodium phosphate buffer (pH 7,0) =

50,0 ml ONPG (8 x 10⁻⁴ M ) = 12,5 mg

For detection of oxidase activity p-Aminodimethylaniline Oxalate = 0,5 g Destilled Water = 50,0 mlNote : To dissolve fully, gently warm the

solution

For immunofluorescence. Solution A : Disodium Phosphate = 1,4 g Distilled water = 100,0 mlSolution B : Sodium dihydrogen phosphate = 1,4 g Distilled water = 100,0 mlNote : Add 84,1 ml of solution A to 15,9 ml of

Solution B. Add 8,5 g of Sodium chloride and q.s. to 1 liter

For detection of coagulase activityNote : Store vials at 2°C to 8°C.

Reconstitute by the addition of 7,5 ml of sterile water

For immunofluorescence Sodium barbital = 6,98 g Sodium chloride = 6,0 g 1 N hydrochloric acid 27,0 ml Distilled water, q.s. to 1000ml

For detedtion of Dnase activity 1% toluidine blue solution = 0,1 ml Distilled water = 99,9 ml

Slowly add 100 ml of 20% aqueous zinc chloride solution to a mixture of 4 g of strach in water. Heat until the strach is dissolved as much as possible and the solution is almost clear. Dilute with water and add 2 g of potassium iodide. Dilute to 1000 ml, filter, and store in an amber bottle

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