The document provides procedures and recipes for making various reagents used in microbiological analysis and testing. It lists the ingredients and instructions for solutions and reagents to detect acetylmethylcarbinol, perform Ames tests, conduct immunofluorescence, detect nitrates, ferric chloride, iodine, catalase activity, indole, and many other microbiology tools. It also provides contact information for the reagent supplier's website, social media, phone number, and email.

1. Prosedur Pembuatan
Reagent Uji Biokimia
Untuk Analisa Mikrobiologi .
www.AlatAlatLaboratorium.com/Blog

3. For detection of acetylmethylcarbinol
Solution A
 Alpha-naphthol = 5,0 g
 Ethanol, Absolute = 95,0 g
Note : 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

4. 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.

5. For ames test
l- Histidine HCl = 0,5 mm
Biotin = 0,5 mm
Distilled Water = 10,0 ml

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

7. 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.

8. 

Ferric Chloride = 10,0 g
Destilled water = 100,0 ml

9. 


Iodine = 1,0 g
Potassium iodide = 2,0 g
Distilled water = 300,0 ml

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

11. For detectin of indole.
p-Dimethylaminobenzaldehyde = 5,0 g
Amly alcohol = 75,0 ml
Hydrochloric Acid (concentrated) = 25,0 ml
Note : Dissolve the p-Dimethylaminobenzaldehyde in
the amly alcohol. Add the hydrochloric Acid.

12. 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

13. 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

14. For microscopic observation of protozoa
Methl cellulose = 10,0 g
Distilled water 90,0 ml

15. For detection of acid
 Methyl red = 0,1 g
 Ethyl alcohol 300,0 ml
 Distilled water = 200,0 ml
Note : Dissolve the methyl red in the 95 % ethyl
alcohol. Dilute to 500 ml with ditilled water

16. For detection of ammonia
 Potassium iodine = 50,0 g
 Distilled water (ammonia-free) = 35,0 ml
Add saturated aqueous solution of mercuric chloride
until a slight precipitate persists.
 Potassium hydroxide (50 % aqueous) = 400,0 ml
Note : 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.

17. 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 ml
Solutiob B, alpha-naphthylamine
 Alpha-naphthylamine = 5,0 g
 Acetic acid, 5 N = 1000,0 ml

18. For enzyme induction
 0,1 M sodium phosphate buffer (pH 7,0) = 50,0 ml
 ONPG (8 x 10⁻⁴ M ) = 12,5 mg

19. For detection of oxidase activity
 p-Aminodimethylaniline Oxalate = 0,5 g
 Destilled Water = 50,0 ml
Note : To dissolve fully, gently warm the solution

20. For immunofluorescence. Solution A :
 Disodium Phosphate = 1,4 g
 Distilled water = 100,0 ml
Solution B :
 Sodium dihydrogen phosphate = 1,4 g
 Distilled water = 100,0 ml
Note : 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

21. For detection of coagulase activity
Note : Store vials at 2°C to 8°C. Reconstitute by the
addition of 7,5 ml of sterile water

22. 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

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

24. 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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