24.4: Materials and Procedures
- Page ID
- 40311
\( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\)
Materials
- Day 1:
- Unknown mixed culture
- 2 TSA plates
- Stains
- Subsequent days: Reagents and stains are available at all times. Media is available by request only.
- TSA plates and slants
- TSB
- Stains
- Physiological media:
- PR Dextrose Broth
- PR Lactose Broth
- PR Sucrose Broth
- MR-VP Broth
- Simmon’s Citrate Slant
- TSI Agar Deep Slant
- Nutrient Gelatin Deep
- Urea Broth
- Starch Agar Plate
- Skim Milk Agar Plate
- EMB Agar Plate
- MacConkey Agar Plate
- Tryptone Agar Plate
- Mueller-Hinton Plate (Novobiocin 30 disk)
- Test Reagents:
- Catalase Reagent-3% Hydrogen Peroxide
- Oxidase Dry slides
- Indole Dry Slides
- Methyl Red
- Barritt’s Reagents A & B
Procedures
Day 1:
- Receive mixed culture, label it, and record #. DO NOT discard this original culture.
- Streak culture onto two TSA plates, incubate.
- Gram Stain mixed culture.
Subsequent Days (activities will be individual in nature, and not everyone will do the same thing on Day 2, Day 3, etc.):
- Subculture ISOLATED colonies only onto a TSA plate, TSA slant, and into a TSB tube, incubate. After incubation observe and record culture characteristics of your plate, slant, and broth. Request media to re-streak cultures if isolation of both bacteria is not achieved.
- Re-Gram stain isolated colonies.
- Request media and run appropriate tests on ISOLATED colonies as needed.
- Record all results.
Contributors and Attributions
Kelly C. Burke (College of the Canyons)