Practice test Laboratory Flashcards

1
Q

In the reverse transcriptase PCR, the cDNA is synthesized by:

a. reverse transcriptase
b. DNA polymerase
c. Oligo dT primers
d. RNAse H

A

DNA polymerase

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2
Q

One of the disadvantages of using a nested PCR is

A

susceptible to contamination

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3
Q

What is the relationship of the fluorescent dye to the single stranded DNA in every PCR cycle?

a. direct relationship
b. inverse relationship
c. no relationship
d. none of these options

A

no relationship

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4
Q

What will happen to the target DNA sequence if the annealing temperature is too high?
1. No hybridization takes place
2. Primers and templates remains dissociated
3. Production of mismatch hybrids
4. Amplification is more likely to occur at nontarget sites in the template molecule

a. 1 and 2
b. 2 and 3
c. 3 and 4
d. 2 and 4
e. 1 and 4

A

1 and 2

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5
Q

What will happen to the target DNA sequence if the annealing temperature is too low?
1. No hybridization takes place
2. Primers and templates remains dissociated
3. Production of mismatch hybrids
4. Amplification is more likely to occur at nontarget sites in the template molecule
a. 1 and 2
b. 2 and 3
c. 3 and 4
d. 2 and 4
e. 1 and 4

A

3 and 4

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6
Q

What would be the effect to the amplicons, if the denaturation temperature is too low?

A

the DNA will not completely denature and amplification efficiency will be low

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7
Q

Which of the following are correct when using a Multiplex PCR?
1. A minimum of three primer sets designed for amplification of different targets are
included in the same PCR reaction.
2. More than one target sequence in a clinical specimen can be amplified in a single
tube.
3. The primers used must be selected carefully to have similar annealing temperatures
should be complementary to each other.
4. The amplicon sizes should be different enough to form distinct bands when visualized
by gel electrophoresis.

a. 1 and 2
b. 1 and 3
c. 2 and 3
d. 2 and 4

A

2 and 4

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8
Q

A type of purification you would use if the sample is for diagnostic analysis.

A

Proteinase K in low salt buffer

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9
Q

Addition of this component can make the DNA pellet stable for long period of time
a. Sterile distilled H2O
b. Heparin
c. EDTA
d. Both sterile distilled water and EDTA

A

Sterile distilled H2O

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10
Q

Any material composed of, containing, or that may contain microbial entities and/or their
harmful products such as toxins and allergens.

A

Biological Material

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11
Q

In a COVID-19 Molecular Biology Laboratory, the main hazard handled is the SARS-COV 2 virus. Which among the following has a potential to cause a laboratory acquired infection
risk?
a. Leak in primary container or viral transport medium
b. DNA extraction inside a Biosafety Cabinet
c. Spillage in the Reagent Preparation Room
d. Non-functional PCR machine

A

Leak in primary container or viral transport medium

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12
Q

Purification of DNA includes removal of proteins, carbohydrates, lipids and cell debris. TRUE OR FALSE

A

TRUE

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13
Q

Ratio of phenol:chloroform:isoamyl alcohol.

A

25:24:1

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14
Q

Staff trained and proficient in techniques is under which basic principle of biosafety?
a. Practices and Procedures
b. Equipment Safety
c. Facility Design and construction
d. Increasing Levels of Protection
e. Material Control
f. None of the above

A

Practices and Procedures

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15
Q

Which among the following about Risk Characterization is NOT TRUE?

a. A hazard cannot pose a risk without a specific situation. Conversely, a situation also does
not represent a risk without a hazard.
b. Both a changing hazard and a changing situation will independently alter the scenario
being assessed, and thus change the risk.
c. The hazard or threat can affect both the likelihood and consequences of a risk, the
situation will usually affect only the likelihood.
d. If an institution finds a particular risk unacceptable, it will either cease the work resulting in
that unacceptable risk, or it will find ways to mitigate that risk to a more acceptable level
e. The actual process of determining the likelihood and consequence of a particular risk
within a Risk Assessment
f. None of the above

A

If an institution finds a particular risk unacceptable, it will either cease the work resulting in
that unacceptable risk, or it will find ways to mitigate that risk to a more acceptable level

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16
Q

Which among the following practices is NOT considered as a security measure designed
to prevent the international release of pathogens.

a. Access control
b. Proper decontamination / disposal of waste material
c. Fences and border lines
d. Personal protective equipment
e. Personnel management
f. None of the above

A

Personnel management

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17
Q

Which of the following evidences must be presented to the Department of Health during
initial monitoring to ensure that the COVID-19 Molecular Laboratory has policies on STAFF
RECRUITMENT, SELECTION, APPOINTMENT AND RESPONSIBILITIES.

a. Written policies and procedures for staff development and training.
b. Proof of training through relevant certificates, memos, written reports, budgetary
allocations.
c. Written policies and procedures on hiring, orientation and promotion of personnel at all
levels
d. Updated organizational chart structured. indicating the names with latest pictures

A
18
Q

It is used to catch nucleotides
I. Glass particles
II. Magnetic beads
III. Silica membrane
IV. Diatomaceous earth

a. I only
b. II and III
c. I, II, IV
d. I, III, IV
e. All of the above

A
19
Q

Which of the following is/are considered as lysing agent?

I. Detergent
II. Zymolyse
III. Lysozyme
IV. Phylase
V. None of the above

a. I only
b. I, II, III
c. I, II, IV
d. V

A

I, II, III

20
Q

Isolation of DNA from whole blood via Ficoll-Directed gradient method what would be the
correct arrangement of the product (top to bottom)

A

Plasma > WBC > Ficoll reagent >
Erythrocytes

21
Q

A 75µL total volume of sample of dsDNA was diluted in 1:5. The diluted sample gave a
reading of 0.451 on a spectrophotometer at OD260. Determine the yield of DNA in the
original sample.

A

8.46 µg DNA

22
Q

Give the DNA concentration in µg/µL.
Dilution: 1:100
Blank reading: 0.680
A260 reading: 1.580
A280 reading: 1.180
a. 0.079 µg/µL
b. 0.79 µg/µL
c. 7.90 µg/µL
d. 79.0 µg/µL
e. 790.0 µg/µL

A

7.90 µg/µL

23
Q

Calculate the yield of DNA in the sample using the following data:
Total volume of DNA sample was 200 mL
The DNA sample was diluted in 1:5
Absorbance at 260 nm of the diluted solution was 0.25

a. 0.0125 µg of DNA
b. 0.125 µg of DNA
c. 1.25 µg of DNA
d. 12.5 µg of DNA
e. 125.0 µg of DNA

A

12.5 µg of DNA

24
Q

Which of the following is true about the length of a primer?

A

If the primers are too short,
they might hybridize to nontarget sites and give undesired amplification products

25
Q

Which of the following troubleshooting procedures must be performed when the annealing
time is too short?

A

use an annealing time of at least 30 seconds.

26
Q

In qPCR, regardless of starting point, all reactions will end at the same, or similar,
concentration. qPCR involves gathering of information during the reaction, when
different reactions are still in the exponential phase.

A
27
Q

Cycle threshold is the number of PCR cycles required to exceed the background
fluorescence. It is also the number of which the fluorescence of the given sample
crosses the threshold value

A
28
Q

Cycle threshold is the number of PCR cycles required to exceed the background
fluorescence. It is also the number of which the fluorescence of the given sample
crosses the threshold value.

A
29
Q

If the denaturation time is too short, the DNA will completely denature and
amplification efficiency will be low. For the initial denaturation, use 3 min to
activate the polymerase and to denature the template during cycling, use 30 sec.

A
30
Q

If the annealing temperature is too high, primers are unable to bind to the
template. The rule of thumb is to use an annealing temperature that is 5°C higher
than the melting point of the primer.

A
31
Q

All of the following are considered pre-analytical standards in Molecular Biology
Diagnostic Laboratories, EXCEPT?

A

Quality Assessment

32
Q

All of the following are principles of Binding of nucleic acid into the membrane except:

A

Number of carbon rings

33
Q

In organic type of purification what part of the solution contains the DNA

a. Upper
b. Lower
c. Middle

A

Upper

34
Q

In Salting out of DNA, what solution you will add in order to further purify the DNA.
a. 100% ethanol
b. Chilled absolute ethanol
c. Saturated NaCl
d. Proteinase K
e. Both a and c

A

Chilled absolute ethanol

35
Q

Is elimination a proper mitigation control for the main hazard (SARS-COV 2 virus) in a
COVID-19 laboratory? Why? Why not?

a. Yes, because elimination is the most efficient mitigation control.
b. Yes, eliminating the main hazard would ensure that the risk of laboratory acquired
infection would be removed.
c. No, substitution is a better mitigation control measure than elimination because you
cannot fully remove handling the virus in the laboratory
d. No, elimination is not possible because the main purpose of the laboratory is to test for the
virus

A

No, elimination is not possible because the main purpose of the laboratory is to test for the
virus

36
Q

Organic type of purification of nucleic acid uses

a. Proteinase K: chloroform: isoamyl alcohol
b. Phenol
c. Absolute alcohol
d. Both a and b

A
37
Q

Other term for Spin column.

A

Solid phase

38
Q

Risk characterization will depend on the likelihood that the risk will happen and the
consequences of the exposure to that risk. Which among the following statements regarding
risk characterization in a COVID-19 molecular laboratory is FALSE?

a. Risk characterization with regards to likelihood will also depend on the frequency of a risk
happening. Because of the high number of samples being submitted to a COVID-19
molecular laboratory, there is a higher possibility of the laboratory to receive viral transport
mediums with leaks. therefore, risk characterization would veer to a higher level of risk.
b. Risk characterization will also depend on the severity of the risk once it occurs. In a
COVID-19 molecular laboratory, exposure of the laboratory staff to the virus while handling it
inside a biosafety cabinet constitutes to a high risk because of the severity of exposure.
c. Risk characterization depicts the mitigation control measure to be applied to a certain
procedure.
d. In a COVID-19 laboratory, the risk of chemical injuries is low. Although exposure to
hazardous chemicals may have severe consequences, it very unlikely to happen because
the molecular laboratory does not handle chemical hazards often.

A

Risk characterization will also depend on the severity of the risk once it occurs. In a
COVID-19 molecular laboratory, exposure of the laboratory staff to the virus while handling it
inside a biosafety cabinet constitutes to a high risk because of the severity of exposure.

39
Q

The following statements regarding equipment maintenance are true, except?

A

Adjustable
and fixed-volume automatic pipettors must be checked for volumetric accuracy and
reproducibility, and recalibrate as necessary before placing in service initially and at specific
defined intervals, at least once every 2 months

40
Q

What would be the sequence in order to get the pure DNA. Cell lysis > affinity via
magnetic beads > elution

A

Cell lysis & affinity via
magnetic beads ; elution

41
Q

Which among the following engineering mitigations is not a recommendation of the
Interim Biosafety Guidelines for Laboratories Handling and Testing SARS-COV 2
(COVID-19) samples.
a. The air must flow from an area of higher contamination to an area of lower contamination.
b. Biosafety Cabinets must be located away from areas where movements that affects
airflow pattern could disrupt the fragile air curtain at the front of the cabinet.
c. There must be a sink with adequate supply for clean water for hand washing located near
the exit door.
d. Access must be restricted to laboratory personnel and support service providers only.

A

he air must flow from an area of higher contamination to an area of lower contamination.

42
Q

Which of the following is used in an Inorganic type of purification?

A

Saturated NaCl