BIOD 171 Lab Notebook 6 Complete Laboratory RecordExam
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HT06 Enzymatic Assays
Objective: To profile bacterial populations based on key enzymatic
reactions and determine growth/metabolic characteristics.
Procedure:
Oxidase test: Qualitative test to determine the presence or absence of
cytochrome c oxidase activity in bacteria.
1. Using a sterile loop pick an isolated colony from
BAP
2. Smear directly onto the reaction area of the
slide.
3. Examine test area for color change within 20
seconds.
a. Oxidase negative = no color change
b. Oxidase positive = purple color
Catalase test: Qualitative test to determine the presence or absence of
catalase, and enzyme used to break down hydrogen peroxide.
1. Using a sterile loop carefully pick colony from
plate
2. Smear colony directly onto the glass slide.
3. Add 2 drops of hydrogen peroxide to each
smear
a. Catalase negative = no bubbles
b. Catalase positive = bubbles
Coagulase test: Qualitative test to determine the presence or absence of
coagulase, an enzyme that plays a role in the formation of
blood clots.
1. Using a sterile loop carefully pick colony from plate
2. Add colony to tub of rabbit plasma, incubate overnight
at 37c
3. (next day) examine samples for precipitants
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a. Coagulase negative = no precipitants (clear and transparent)
b. Coagulase positive = presence of fibrin aggregates (clots)
i. Small specs in the picture are clots
ii. The milky cloudiness is a precipitate forming
Lipase Test: Qualitative test to determine the presence or absence of lipase,
and enzyme that hydrolyzes triglycerides (fatty acids/lipids).
1. Using a sterile loop unknown culture onto spirit blue
plates
2. Streak colony across plate, incubate overnight at 37c
3. (Next day) examine sample for precipitants
a. Lipase negative = no change
b. Lipase positive = reduction in color (zone of
clearance)
i. Begun to hydrolyze the fatty acids, which changed the pH
and the acidity of the media.
ii. Similar to beta hemolysis on blood agar plates
Wet Lab: Using E coli and pseudomonas (both gram neg), which on EMB
plate. E coli fermented lactose and turned green, whereas pseudomonas did
not.
Oxidase Test:
1. We have three oxidase test strips, make a note, 0 have negative
control, just water will be added to ensure a false
positive doesn’t occur
2. Write E coli and pseudomonas on the paper
3. Add a small drop of water to moisten the strip
4. Harvest from the 4 quadrant plate using sterile loop
5. Drag/mis bacteria onto strip, gentle swirls
6. Now go to pseudomonas strain, repeat, smear into
test strip.
Catalase test:
1. Go back to 4 quadrant plate
2. Two are gram positive (staph aureus and
streptococcus)
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Notebook 6 Complete Laboratory Record and Study Guide With Solution.pdf
and Study
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BIOD
Guide
171Study
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Lab Notebook
Guide
Solution
With
6 Solution
Complete Laboratory Record and Study Guide With Solution.pdf
Lab Notebook
HT06 Enzymatic Assays
Objective: To profile bacterial populations based on key enzymatic
reactions and determine growth/metabolic characteristics.
Procedure:
Oxidase test: Qualitative test to determine the presence or absence of
cytochrome c oxidase activity in bacteria.
1. Using a sterile loop pick an isolated colony from
BAP
2. Smear directly onto the reaction area of the
slide.
3. Examine test area for color change within 20
seconds.
a. Oxidase negative = no color change
b. Oxidase positive = purple color
Catalase test: Qualitative test to determine the presence or absence of
catalase, and enzyme used to break down hydrogen peroxide.
1. Using a sterile loop carefully pick colony from
plate
2. Smear colony directly onto the glass slide.
3. Add 2 drops of hydrogen peroxide to each
smear
a. Catalase negative = no bubbles
b. Catalase positive = bubbles
Coagulase test: Qualitative test to determine the presence or absence of
coagulase, an enzyme that plays a role in the formation of
blood clots.
1. Using a sterile loop carefully pick colony from plate
2. Add colony to tub of rabbit plasma, incubate overnight
at 37c
3. (next day) examine samples for precipitants
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/2026
Notebook 6 Complete Laboratory Record and Study Guide With Solution.pdf
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Guide
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a. Coagulase negative = no precipitants (clear and transparent)
b. Coagulase positive = presence of fibrin aggregates (clots)
i. Small specs in the picture are clots
ii. The milky cloudiness is a precipitate forming
Lipase Test: Qualitative test to determine the presence or absence of lipase,
and enzyme that hydrolyzes triglycerides (fatty acids/lipids).
1. Using a sterile loop unknown culture onto spirit blue
plates
2. Streak colony across plate, incubate overnight at 37c
3. (Next day) examine sample for precipitants
a. Lipase negative = no change
b. Lipase positive = reduction in color (zone of
clearance)
i. Begun to hydrolyze the fatty acids, which changed the pH
and the acidity of the media.
ii. Similar to beta hemolysis on blood agar plates
Wet Lab: Using E coli and pseudomonas (both gram neg), which on EMB
plate. E coli fermented lactose and turned green, whereas pseudomonas did
not.
Oxidase Test:
1. We have three oxidase test strips, make a note, 0 have negative
control, just water will be added to ensure a false
positive doesn’t occur
2. Write E coli and pseudomonas on the paper
3. Add a small drop of water to moisten the strip
4. Harvest from the 4 quadrant plate using sterile loop
5. Drag/mis bacteria onto strip, gentle swirls
6. Now go to pseudomonas strain, repeat, smear into
test strip.
Catalase test:
1. Go back to 4 quadrant plate
2. Two are gram positive (staph aureus and
streptococcus)
This study source was downloaded by 1827175 from cliffsnotes.com on 01-08-2026 18:31:09 GMT -06:00
Page 2 of 6
https://www.cliffsnotes.com//study-notes/24920345 BIOD 171 Lab
/2026
Notebook 6 Complete Laboratory Record and Study Guide With Solution.pdf
