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Techniques for Microbiology


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Table of Contents

Unit I - Safety first!

Safety in the microbiology laboratory

Universal precautions

Unit II - Culturing bacteria and aseptic techniques

Preparing culture media


The autoclave

Dry heat

Aseptic transfer of bacteria

Adjusting the gas burner

Preparing bacterial cultures in broth tubes and on agar slants

Transfer colony from plate to broth and to agar slant

Isolation of bacteria

Preparing a pour plate

Preparing a streak plate

Preparing a spread plate

Characteristic features of bacterial growth in broth and on agar

Maintenance and storage of stock cultures

Culturing anaerobic bacteria

Unit III - Visualizing bacteria

Effective use and responsible care of the light microscope

Measuring microscopic cells

Preparing a hanging-drop slide

Preparing a bacterial smear

Preparing a simple stain

Preparing a Gram stain

Preparing an acid-fast stain

Preparing a negative stain

Preparing a capsule stain

Preparing an endospore stain

Unit IV - Enzyme-based tests for the identification of bacteria

Hydrolytic (digestive) enzymes

Starch hydrolysis

Casein hydrolysis

Gelatin hydrolysis

Lipid hydrolysis (tributyrin and spirit blue agar)

DNA hydrolysis

Utilization of carbohydrates

Fermentation of carbohydrates (Durham tube)

Methyl red test (mixed fermentation)

Voges-Proskauer test (butanediol fermentation)

Citrate utilization

Oxidation-fermentation (O-F) glucose test

Degradation of amino acids

Indole test

H2 S production

Phenylalanine deamination

Decarboxylase test

Respiration tests

Catalase test

Oxidase test

Nitrate reduction

Miscellaneous tests

Triple sugar iron (TSI) test

Urea hydrolysis

Litmus milk reactions

Motility assay

Coagulase production

Selective and/or differential media

Blood agar

Eosin methylene blue (EMB) agar

Mannitol-salt agar

MacConkey agar

Phenylethyl alcohol (PEA) agar

Unit V - Counting microbes

Direct microscopic counting with the Petroff-Hausser chamber

Preparing a standard plate count of bacteria

Using Turbidemetry to estimate cell density

Plaque assay for determining bacteriophage titer

Unit VI - Measuring effectiveness of antibacterial chemicals

Kirby-Bauer method for sensitivity of bacteria to antibacterial


Evaluating antibacterial chemicals: The disk-diffusion


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