1. Identification of Unknown Bacteria: individual assignment
Lab 12 Goals and Objectives:
Identification of Unknown Bacteria: individual assignment
Use your data from the 12 possible unknowns for comparison and your Dichotomous Key to solve for your unknown based on the results you collect.
From single colonies:
Inoculate one MacConkey Agar (streak for isolation)
Inoculate one Nutrient Gelatin Deep (stab)
Make new streak for isolation on BHIA
Make Gram Stain slides (at least 2 incase you drop one)
record Gram reaction, size of cells, shape of cells, any
arrangements you observe
Test for catalase production on a slide

2. Unknown Identification
Day 1: Each person sign out one unknown
Make streak for isolation on BHIA
Put your name on original slant and return
Day 2: Gram stain
Catalase test
Inoculate MacConkey Agar
Inoculate Gelatin Deep
Day 3: Based on Gram, Gelatinase, Catalase,
Lactose, set up assays to solve:
A. If Staph: MSA and Coagulase
B. If Strep/Entero: BAP and Bacitracin
C. If Gram Neg: SIM, Citrate, MR, Urea

3. All 12 Possible Unknowns
Gram Positive
Gram Negative
Gelatinase +
Gelatinase -
Gelatinase -
Gelatinase +
Catalase +
Catalase -
Lactose +
Lactose -
Mannitol + OR
Coagulase +
Mannitol - OR
Coagulase -
 Hemolytic OR
Bacitracin R
 Hemolytic OR
Bacitracin S
Indole + OR
Citrate -
Indole - OR
Citrate +
H2S + OR
Motile
H2S - OR
Nonmotile
Urea + OR
MR +
VP -
Urea - OR
MR -
VP +
Urea +
Urea -
Staphylococcus epidermidis
Pseudomonas aeruginosa
Staphylococcus aureus
Enterococcus faecalis
Streptococcus pyogenes
Klebsiella pneumoniae
Salmonella typhimurium
Enterobacter cloacae
Bacillus subtilis
Escherichia coli
Proteus vulgaris
Shigella flexneri

4. Identification of Unknown Bacteria: individual assignment
Lab 12 Goals and Objectives:
Identification of Unknown Bacteria: individual assignment
Use your data from the 12 possible unknowns for comparison and your Dichotomous Key to solve for your unknown based on the results you collect.
From single colonies:
Inoculate one MacConkey Agar (streak for isolation)
Inoculate one Nutrient Gelatin Deep (stab)
Make new streak for isolation on BHIA
Make Gram Stain slides (at least 2 incase you drop one)
record Gram reaction, size of cells, shape of cells, any
arrangements you observe
Test for catalase production on a slide