,Microbiology Laboratory Final Exam Study
Guide – Complete Lab Techniques,
Experiments & Practice Questions.
there will be three parts to the final:
-1st part: we will be required to perform a test we have done since the midterm, interpret the
results of the test, and answer questions **will not be oxidase since we didn't do that
-2nd part: we will have to answer questions about materials (broths, plates, etc) on the bench
top in front of us. Media like Snyder, nitrate broth, EMB, etc. Understand why we used this
media.
-3rd part: we will have to answer a variety of questions about lab techniques 1-8, experiments
1-13, and the microbiome project
*names of organisms?
*exp 8-13 are important
**know how to do nitrate reduction bc its like 4 parts
the practical
-Catalase, oxidase, indole, Methyl red, Vogues-Proskauer, or Nitrate reduction test
midterm quizlet goes up to lab technique 4, experiment 4, and quiz 4
review lab technique 5-8, plating, use of a microscope, use of pipettes, and serial dilutions
indole test
-media used
- enzyme ______ breaks down ______ into ______, ______, and ______
-Kovac's reagent:
**check for indole production by adding a dropper full of Kozak's reagent--reacts with indole
to form red layer at top of broth (+ result)
-1% tryptone broth
- tryptophanase
,-tryptophan
-indole, pyruvic acid, and ammonia
methyl red test
-Some organisms (E. coli) use mixed acid fermentation to ferment glucose and produce
several acids, pH drops
-Add a dropper full pH indicator methyl red to one tube of MRVP broth
-Methyl red turns red below pH 4.2 (+ result)
Vogues-Proskauer test
-Some organisms ferment glucose by producing acetyl methyl carbinol (acetone) from
pyruvate
-follow up on MRVP broth
1) Add a dropper full Alpha Napthol and 40% KOH
2) Vortex
3) let sit undisturbed for 20 min. These reagents will react with acetone to produce a wine-red
color (+ result) in the upper part of the broth where oxygen is present
Citrate test
- Inoculate Simmons citrate agar slant
- _______ is only carbon source
-___________ is only nitrogen source
-_______is pH indicator (green at pH 7), blue above pH 7
- some microorganisms can utilize ____ as the sole carbon source
- bacteria that can do this can also convert ammonia phosphate to ________ and _________
- Sodium citrate
-Ammonium Phosphate
-Bromothymol blue is pH indicator
-citrate
- ammonia
-ammonia hydroxide
exp 5 quiz
pic
pic
, pic
pic
just add all the dots up (17) and multiply by 10^4
exp 5: motility
________ are used by many microorganisms for locomotion. Although flagella may be found
among all three domains of life, the structure of the flagella are quite different. The
eukaryotic flagellum is made of_______ (made of the protein tubulin) driven by dynein motor
proteins, which move eukaryotic flagella back and forth. _______ is a single-celled eukaryotic
green alga with a pair of flagella at one end of the cell each whipping back and forth to move
the cell. The prokaryotic flagellum is far more complex.
Bacterial and archaeal flagella appear similar in structure superficially, but on closer
examination there are subtle structural differences such as the width of the flagella as well as
how the flagella use energy. Bacterial flagella are anchored by a complex of proteins in the
cell envelope called the ______ or motor. A curved hook connects the basal body to the
extracellular component of the flagellum, the filament. The filament is made of _______.
-Flagella
- microtubules
-Chlamydomonas reinhardtii
-basal body
-flagellin proteins
The current model for the movement of bacterial flagella is by a “rotary engine” (or “rotary
motor”) in which a proton gradient (Na+, K+, and Rb+ can also be used) provides the energy
to rotate the flagellum. The force to spin the motor proteins can be obtained when protons
from the periplasm flow back into the cell through ______, which are located adjacent to the
motor.
The number and attachment of bacterial flagella can vary among species and is characteristic
of a given species. When one or more flagella are attached at the “ends” of the cells, the
attachment is considered _____. Polar attachment can take many forms. For example, a single
flagellum may be attached at one end of a cell in a _______fashion. When a “tuft” of flagella
are attached at one or both ends, the flagellar arrangement is called _______ A bacterium
that has one flagella at each end of the cell is said to have ______ flagella. In contrast to the
Guide – Complete Lab Techniques,
Experiments & Practice Questions.
there will be three parts to the final:
-1st part: we will be required to perform a test we have done since the midterm, interpret the
results of the test, and answer questions **will not be oxidase since we didn't do that
-2nd part: we will have to answer questions about materials (broths, plates, etc) on the bench
top in front of us. Media like Snyder, nitrate broth, EMB, etc. Understand why we used this
media.
-3rd part: we will have to answer a variety of questions about lab techniques 1-8, experiments
1-13, and the microbiome project
*names of organisms?
*exp 8-13 are important
**know how to do nitrate reduction bc its like 4 parts
the practical
-Catalase, oxidase, indole, Methyl red, Vogues-Proskauer, or Nitrate reduction test
midterm quizlet goes up to lab technique 4, experiment 4, and quiz 4
review lab technique 5-8, plating, use of a microscope, use of pipettes, and serial dilutions
indole test
-media used
- enzyme ______ breaks down ______ into ______, ______, and ______
-Kovac's reagent:
**check for indole production by adding a dropper full of Kozak's reagent--reacts with indole
to form red layer at top of broth (+ result)
-1% tryptone broth
- tryptophanase
,-tryptophan
-indole, pyruvic acid, and ammonia
methyl red test
-Some organisms (E. coli) use mixed acid fermentation to ferment glucose and produce
several acids, pH drops
-Add a dropper full pH indicator methyl red to one tube of MRVP broth
-Methyl red turns red below pH 4.2 (+ result)
Vogues-Proskauer test
-Some organisms ferment glucose by producing acetyl methyl carbinol (acetone) from
pyruvate
-follow up on MRVP broth
1) Add a dropper full Alpha Napthol and 40% KOH
2) Vortex
3) let sit undisturbed for 20 min. These reagents will react with acetone to produce a wine-red
color (+ result) in the upper part of the broth where oxygen is present
Citrate test
- Inoculate Simmons citrate agar slant
- _______ is only carbon source
-___________ is only nitrogen source
-_______is pH indicator (green at pH 7), blue above pH 7
- some microorganisms can utilize ____ as the sole carbon source
- bacteria that can do this can also convert ammonia phosphate to ________ and _________
- Sodium citrate
-Ammonium Phosphate
-Bromothymol blue is pH indicator
-citrate
- ammonia
-ammonia hydroxide
exp 5 quiz
pic
pic
, pic
pic
just add all the dots up (17) and multiply by 10^4
exp 5: motility
________ are used by many microorganisms for locomotion. Although flagella may be found
among all three domains of life, the structure of the flagella are quite different. The
eukaryotic flagellum is made of_______ (made of the protein tubulin) driven by dynein motor
proteins, which move eukaryotic flagella back and forth. _______ is a single-celled eukaryotic
green alga with a pair of flagella at one end of the cell each whipping back and forth to move
the cell. The prokaryotic flagellum is far more complex.
Bacterial and archaeal flagella appear similar in structure superficially, but on closer
examination there are subtle structural differences such as the width of the flagella as well as
how the flagella use energy. Bacterial flagella are anchored by a complex of proteins in the
cell envelope called the ______ or motor. A curved hook connects the basal body to the
extracellular component of the flagellum, the filament. The filament is made of _______.
-Flagella
- microtubules
-Chlamydomonas reinhardtii
-basal body
-flagellin proteins
The current model for the movement of bacterial flagella is by a “rotary engine” (or “rotary
motor”) in which a proton gradient (Na+, K+, and Rb+ can also be used) provides the energy
to rotate the flagellum. The force to spin the motor proteins can be obtained when protons
from the periplasm flow back into the cell through ______, which are located adjacent to the
motor.
The number and attachment of bacterial flagella can vary among species and is characteristic
of a given species. When one or more flagella are attached at the “ends” of the cells, the
attachment is considered _____. Polar attachment can take many forms. For example, a single
flagellum may be attached at one end of a cell in a _______fashion. When a “tuft” of flagella
are attached at one or both ends, the flagellar arrangement is called _______ A bacterium
that has one flagella at each end of the cell is said to have ______ flagella. In contrast to the
