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Test Bank For Microbiology A Laboratory Manual 9th Edition By Cappuccino APPENDIX 2 Biochemical Test Reagents Barritt’s reagent, for detection of acetylmethylcarbinol Solution A Not

Test Bank For Microbiology A Laboratory Manual

9th Edition By Cappuccino

APPENDIX 2

Biochemical Test Reagents

Barritt’s reagent, for detection of acetylmethylcarbinol

Solution A

Note: Dissolve the alpha-naphthol in the ethanol with constant stirring

Solution B

Note: Dissolve the potassium hydroxide in 75 ml of distilled 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

Biotin-histidine solution, for Ames test

Buffered glycerol (pH 7.2), for immunofluorescence

Phosphate buffered saline 10.0 ml

Diphenylamine reagent, for detection of nitrates

Dissolve 0.7 g diphenylamine in a mixture of 60 ml concentrated sulfuric acid and 28.8 ml of distilled water Cool and slowly add 11.3 ml of concentrated hydrochloric acid Allow to stand for 12 hours Sedimentation indicates that the reagent is saturated

Ferric chloride reagent

Gram’s iodine, for detection of starch

As in Gram stain

Hydrogen peroxide, 3%, for detection

of catalase activity

Note: Refrigerate when not in use

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Kovac’s reagent, for detection of indole

p-Dimethylaminobenzaldehyde 5.0 g

Hydrochloric acid (concentrated) 25.0 ml

Note: Dissolve the p-dimethylaminobenzaldehyde in the amyl alcohol Add the hydrochloric acid

McFarland Barium Sulfate Standards, for STAPH-IDENT procedure

Prepare 1% aqueous barium chloride and 1% sulfuric acid solutions Using the table below, add the amounts of barium chloride and sulfuric acid to clean 15-  150-mm screw-cap test tubes Label the tubes 1 through 10

Preparation of MacFarland Standards

Corresponding approximate

Methyl cellulose, for microscopic observation of protozoa

Methyl red solution, for detection of acid

Note: Dissolve the methyl red in the 95% ethyl alcohol Dilute to 500 ml with distilled water

Nessler’s reagent, for detection of ammonia

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

Nitrate test solution, for detection of nitrites

Solution A, Sulfanilic acid

Acetic acid, 5 N: 1 part glacial

acetic acid to 2.5 parts

Solution B, Alpha-naphthylamine

Acetic acid, 5 N 1000.0 ml

Orthonitrophenyl-ß-D-galactoside (ONPG), for enzyme induction

0.1 M sodium phosphate

p-Aminodimethylaniline oxalate, for detection of oxidase activity

p-Aminodimethylaniline oxalate 0.5 g

Note: To dissolve fully, gently warm the solution

Phosphate-buffered saline, 1% (pH 7.2–7.4), for immunofluorescence

Solution A

Solution B

Sodium dihydrogen phosphate 1.4 g

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

Rabbit plasma, 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

Sodium barbital buffer, for immunofluorescence

1 N hydrochloric acid 27.0 ml

Distilled water, q.s to 1000 ml

Toluidine blue solution, 0.1%, for detection of DNase activity

1% toluidine blue solution 0.1 ml

Tris-acetate buffer 10 

Note: Add ingredients to 1-liter volumetric flask and then add water to volume

Tris-acetate buffer 1 

Tris-acetate buffer 10

Note: Buffer can be stored indefinitely at room temperature

Tris-borate buffer 5 

Tris-borate buffer 1  working solution

Tris-borate buffer 5

Trommsdorf’s reagent, for detection of nitrite

Slowly add 100 ml of 20% aqueous zinc chloride solution to a mixture of 4 g of starch in water Heat until the starch

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