Lab exam 2: objectives Flashcards
What is the name of the pink oxygen indicator?
methylene pink
resazurin
methylene blue
phenol red
resazurin
How should a microbe ONLY growing at the very top of a thioglycollate tube be decribed with respect to oxygen?
Facultative anaerobe
Obligate anaerobe
Obligate aerobe
Obligate aerobe
How should a microbe growing in the pink and below the pink in a thioglycollate tube be classified with respect to oxygen?
Obligate anaerobe
Facultative anaerobe
Aerobe
Cannot be determined
Facultative anaerobe
How should a microbe growing ONLY without oxygen be classified with respect to oxygen?
Aerobe
Facultative anaerobe
Obligate anaerobe
Obligate anaerobe
A microbe that grows with and without oxygen is a(n):
obligate aerobe
obligate anaerobe
facultative anaerobe
facultative anaerobe
An organism that grows well in the incubator and slightly less in the GasPak is a(n)
Facultative anaerobe
Obligate anaerobe
Obligate aerobe
Cannot be determined
Facultative anaerobe
What is the name of the enzyme that hydrolyzes casein?
amylase
casease
lipase
casease
How will you know if the bacteria hydrolyze casein?
A clear zone surrounding the growth will be visible following incubation.
A cloudy zone surrounding the growth will be visible following incubation
The area surrounding the growth will turn dark purple.
A clear zone surrounding the growth will be visible following incubation.
Starch is hydrolyzed using:
intracellular enzymes
extracellular enzymes
extracellular enzymes
Casein is a:
nucleic acid
protein
carbohydrate
protein
The products of protein hydrolysis are:
monosaccharides
amino acids
glycerol and fatty acids
amino acids
A product of starch hydrolysis is:
monosaccharide
amino acid
glycerol
monosaccharide
PCR is an abbreviation for
Polymerase carboxyl reaction
Polymerase chain reaction
Polymer chain reaction
Polymerase chain reaction
PCR is like a molecular xerox machine for DNA. The process is designed to make multiple copies (amplify) of a desired gene or other short DNA fragment.
True
False
True
PCR rapidly _______ the amount of DNA in a sample. This decreases the need for making cultures or carrying out complex purification techniques. Once multiple copies of DNA are made, they can be used in various ways, ranging from research to _________________.
Decreases….diagnostics and forensics
Increases….culturing bacteria and fungi
Increases…diagnostics and forensics
Increases…diagnostics and forensics
Since DNA is tiny and colorless, the resulting PCR products cannot be visualized in the reaction tube. Agarose gel electrophoresis will allow visualization of the DNA as bands in the gel.
True
False
True
PCR makes it possible to amplify a target DNA from a few copies to at least_______ of copies in a few hours.
Hundreds
Thousands
Millions
Millions
________________are short single-stranded DNA strands of a known sequence that indicate where DNA amplification will begin.
DNA polymerase
Primers
Nucleotides
Primers
Taq polymerase is an example of a DNA polymerase used in PCR. This enzyme works at
Low temperatures
Relatively high temperatures
High salt concentration
Relatively high temperatures
The machine used in PCR is called the
Thermal cycler
PCR prepper
Thermal vent polymerase
Thermal cycler
In the _________ step, primers bind to the test DNA. This occurs around 45-65 degrees C. The temperature allows the primers to complementary base pair to their target sequences on opposite strands of the template DNA.
Denaturation
Annealing
Extension
Annealing
In the ________ step, target DNA is heated to 92-96 degrees C so the double stranded DNA will separate into 2 strands. The heat breaks the hydrogen bonds holding the double-stranded DNA together.
Denaturation
Priming
Annealing
Denaturation
The _______ step occurs around 72 degrees C. The DNA polymerase uses the raw materials in the form of nucleotides to extend the molecule and produce 2 complete strands of DNA.
Denaturation
Priming
Extension
Extension
Which of the following ingredients must be in a PCR reaction?
DNA template
Primers
Nucleotides
DNA polymerase with the appropriate buffer
All of the above
All of the above
Your unknown is a member of the Enterobacteriaceae family (enterics). See index or research online if needed. Therefore, it will be
Gram-positive
Gram-negative
Gram-negative
The description of your Gram-positive colony may be taken from your______plates. Read PEA and Mac labs.
PEA or TSA (nutrient agar)
MacConkey or TSA (nutrient agar)
PEA or TSA (nutrient agar)
When you perform your Gram stain from your MacConkey plate, you should see
Gram-positive cocci
Gram-negative rods or coccobacilli
Gram-positive rods or coccobacilli
Gram-negative rods or coccobacilli
Please see your lab schedule for this question. We will be further purifying the Gram-negative unknown away from the Gram-positive contaminant. Therefore, we will pick a colony of the Gram-negative microbe and
grow it in the Gas Pak
grow it on PEA
streak for isolation again onto a TSA plate
streak for isolation again onto a TSA plate
Please refer to your lab schedule. Your unknown may be ______________. Select all of the correct choices.
Serratia marcescens
Enterococcus faecalis
Escherichia coli
Bacillus subtilis
Klebsiella pnuemoniae
Proteus vulgaris
Staphylococcus aureus
Serratia marcescens
Escherichia coli
Klebsiella pnuemoniae
Proteus vulgaris
Please read PEA and MAC labs. The description of your Gram-negative colony may be taken from your______plates.
PEA or TSA (nutrient agar)
MacConkey or TSA (nutrient agar)
MacConkey or TSA (nutrient agar)
With regard to the phenol red broth, what is the function of the Durham tube?
collect gas
trap aerobes
pH indicator
collect gas
SIM medium is inoculated with a _________ because we can check for _______.
needle….oxygen use
loop….turbidity
needle…motility
needle…motility
SIM medium may be used to:
differentiate pathogens that cause neurological disorders
differentiate Enterobacteriaceae
differentiate respiratory pathogens
differentiate Enterobacteriaceae
The sole carbon source in citrate medium is______. This test will help you distinguish among the unknowns.
citrate
glucose
phosphate
citrate
All of the tests we are setting up for this lab will help differentiate your unknown, so it is important to inoculate carefully.
False
True
True
You will inoculate your unknown into the Enteropluri tube. This test will allow us to perform multiple tests in a single tube. There are ______ individual chambers.
2
6
12
12
A(n) _______________________ is the material used to separate DNA fragments.
Reaction mix
Buffer
Agarose gel
Agarose gel
The _________are where you put the DNA in a gel at the top.
Dips
Wells
Gels
Wells
________bands of DNA will migrate the farthest in the agarose gel.
Heavier
Lighter
Lighter
TAE or TBE are types of ___________used during the electrophoresis process.
Enzyme
Buffer
Gel
Buffer
DNA is negatively charged and will run toward the positive pole in gel electrophoresis.
True
False
True
A phenol red broth after incubation appeared yellow with a bubble in the Durharm tube. How would you record this tube?
no reaction
acid without gas
acid and gas
protein degradation
acid and gas
In the phenol red broths, if an organism released an acidic product, the pH indicator turns:
Pink
Yellow
Red
Blue
Yellow - pH below 6.8 (acidic)
Red - neutral
Pink - pH above 7.4 (alkaline)
The black color in a SIM tube indicates the bacteria produced a substance called___________.
FeSO4
indole
H2S
FeS
H2S
With regard to the previous question on SIM medium, the actual chemical representing the black precipitate is ___________.
indole
H2S
FeSO4
FeS
FeS
A bright red color is observed after an indole test is performed from SIM medium. Is the indole test positive or negative for this organism?
Positive
Negative
Positive
Would the indole test work if tryptophan was accidentally left out of this media?
Yes
No
No
Name the reagent added to perform the indole test.
Phenol Red
Methyl Red
Ferric chloride
Kovak’s reagent
Kovak’s reagent
An organism that utilizes citrate will appear:
as no color change with no growth
blue or no color change, but with growth
blue or no color change, but with growth
MacConkey agar is:
selective and differential
enriched and differential
selective only
differential only
selective and differential
The use of the streak plate technique is to:
obtain isolation of individual species from a mixed sample.
How many times will you dip into your original culture when performing a streak plate?
only once, to streak the first quadrant.
The gram stain is a:
Simple stain
Differential stain
Negative stain
Differential stain
Describe 2 methods for providing anaerobic growth conditions:
Fluid thioglycolate broth (FTB)
Anaerobic jar (gaspak)
Know which components of the medium reduce oxygen to water:
Sodium thioglycolate
L-cystine
Sodium thioglycolate
L-cystine
Interpret the results of a citrate test:
Blue
no color change (growth)
no color change (no growth)
Blue - Positive (+ citrate permease is present)
No color change (growth) - Positive
No color change (no growth) - Negative
Define Coliforms:
most are normal inhabitants of the human intestinal tract; their presence in the environment may be evidence of fecal contamination.
a subgroup of Enterobacteriaceae that produce acid and gas from lactose fermentation.
Define Enterics:
Pathogenic bacteria of bacterial gastroenteritis.
bacteria of the intestines, and may refer to gut flora, which are always present and usually harmless.
Coliforms are normal gram-_______ inhabitants of the human ________.
positive…skin
negative…skin
positive…intestinal tract
negative…intestinal tract
negative…intestinal tract
Match the types of arrangements:
Diplo- Chains
Strepto- Irregular clumps, grapes
Staphylo- Pairs
Diplo - pairs
Strepto- chains
Staphylo - irregular clumps, grapes
Phenylethyl alcohol agar (PEA) is mostly selective for _________ organisms. This is because PEA inhibits the growth of many Gram-negative while only some Gram-positive organisms. PEA is _________________.
Gram-positive….selective and differential
Gram-positive….selective only
Gram-negative…selective and differential
Gram-negative…selective only
Gram-positive….selective only
A hot pink color was observed on colonies growing on MacConkey agar. These organisms are capable of _______________ fermentation.
glucose
lactose
sucrose
galactose