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[OS 201] 3rd Exam > Trans - Utility of Molecular Tools in Health Care and Biological Research > Flashcards

Trans - Utility of Molecular Tools in Health Care and Biological Research Flashcards

1
Q

discovered DNA double helix

A

James Watson, Francis Crick

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2
Q

BRCA 1 & 2

A

transcriptional molecules correlated with breast cancer (5% risk with BRCA 1; more risk with BRCA 2)

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3
Q

physical manifestation of genetic variation

A

anatomic variation

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4
Q

biochemical manifestation of genetic variation

A

physiologic

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5
Q

main genomic variations [3]

A

[1] single base pair change
[2] insertions, deletions
[3] structural rearrangements (ex. translocation)

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6
Q

epigenetic mechanism of genetic variation

A

[1] methylation of DNA

[2] histone packing

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7
Q

epigenetic mechanism affected by: [5]

A 
[1] development (in utero, childhood)
[2] environmental chemicals
[3] drugs
[4] aging
[5] diet
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8
Q

methods of nucleic acid extraction

A

[1] organic-based extraction

[2] silica-based extraction

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9
Q

methods of nucleic acid amplification

A

[1] polymerase chain reaction

[2] cloning

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10
Q

post-amplification methods [4]

A

[1] direct assessment
[2] electrophoresis
[3] hybridization assay
[4] sequencing

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11
Q

when will you use organic methods of nucleic acid extraction?

A

when you want a greater quantity of nucleic acid

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12
Q

when will you use silica column method of nucleic acid extraction?

A

when you want greater purity of the sample

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13
Q

principle of nucleic acid extraction by organic solvent

A

[1] aqueous chloroform solution separates hydrophilic DNA from other components
[2] when alcohol is added, DNA precipitates at inferface –> easily collected

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14
Q

2 main organic solvents used in nucleic acid extraction

A

[1] chloroform

[2] alcohol

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15
Q

requirements of PCR [4]

A

[1] DNA template
[2] DNA polymerase
[3] DNA primers
[4] ions and pH buffers

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16
Q

PCR - temperature to activate DNA polymerase

A

94-96 C

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17
Q

PCR - denaturation step temperature

A

94-96C

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18
Q

PCR - annealing step - temperature and purpose

A

50-65 C to allow primers to bind to complementary strands

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19
Q

PCR - extension step - temperature and purpose

A

72 C to allow elongation

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20
Q

PCR - final extension step - temperature and purpose

A

70-74 C to ensure remaining single stranded DNA is fully extended

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21
Q

PCR - final hold - temperature and purpose

A

4-15 C for short-term storage

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22
Q

DNA polymerase used in PCR? why?

A

Taq polymerase - heat resistant polymerase from Thermus aquaticus

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23
Q

electrophoresis - principle

A

separation of DNA segments by size and charge via migration through gels driven by electrical charges

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24
Q

electrophoresis - purpose

A

to obtain a certain DNA segment of interest

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25
Q

electrophoresis - direction of flow? why?

A

towards the + terminal, because DNA is negatively charged

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26
Q

electrophoresis - rate of flow is affected by what factors?

A

rate of flow determined by size (larger = slower) and charge (more positive = slower)

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27
Q

LAMP - meaning

A

Loop Mediated Isothermal Amplification of DNA

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28
Q

LAMP - principle

A

PCR conducted at only one temperature

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28
Q

LAMP - principle

A

PCR conducted at only one temperature

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29
Q

LAMP - microorganism involved

A

bacillus stereos thermophiles (BST)

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29
Q

LAMP - microorganism involved

A

bacillus stereos thermophiles (BST)

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30
Q

LAMP - temperature

A

65 C

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30
Q

LAMP - temperature

A

65 C

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31
Q

LAMP - major disadvantages [2]

A

[1] takes too long

[2] nonspecificity

31
Q

LAMP - major disadvantages [2]

A

[1] takes too long

[2] nonspecificity

32
Q

LAMP - major advantages [3]

A

[1] cheaper than PCR
[2] easier to do
[3[ may be done in remote areas

32
Q

LAMP - major advantages [3]

A

[1] cheaper than PCR
[2] easier to do
[3[ may be done in remote areas

33
Q

hybridization assay - principle

A

probes are used to determine if target sequence is present or not in a given DNA sample

33
Q

hybridization assay - principle

A

probes are used to determine if target sequence is present or not in a given DNA sample

34
Q

pharmacogenesis

A

concerned with drug metabolism

34
Q

pharmacogenesis

A

concerned with drug metabolism

35
Q

pharmacogenesis - mainly controlled by what factor

A

60 cytochrome P450

35
Q

pharmacogenesis - mainly controlled by what factor

A

60 cytochrome P450

36
Q

pharmacogenesis - ways of altering drugs

A

[1] drug to metabolite (which may be toxic)

[2] prodrug (which may be toxic) to drug

36
Q

pharmacogenesis - ways of altering drugs

A

[1] drug to metabolite (which may be toxic)

[2] prodrug (which may be toxic) to drug

37
Q

warfarin

A

effective anticoagulant

37
Q

warfarin

A

effective anticoagulant

38
Q

involved in warfarin metabolism [2]

A

[1] CYS269 - metabolizes warfarin

[2] VKORC1 - prevents bleeding

38
Q

involved in warfarin metabolism [2]

A

[1] CYS269 - metabolizes warfarin

[2] VKORC1 - prevents bleeding

39
Q

microarray technology - principle

A

information on more than a million unique sequences and variances may be stored in a single chip

39
Q

microarray technology - principle

A

information on more than a million unique sequences and variances may be stored in a single chip

40
Q

steps in microarray processing [4]

A

[1] creation of array of oligonucleotides on a chip
[2] hybridization of fluorescently labeled DNA/RNA
[3] scanning of the array
[4] computational processing of raw image

40
Q

steps in microarray processing [4]

A

[1] creation of array of oligonucleotides on a chip
[2] hybridization of fluorescently labeled DNA/RNA
[3] scanning of the array
[4] computational processing of raw image

41
Q

microarray technology - importance

A

ability to visually determine which variations of DNA will cause expression of certain traits

41
Q

microarray technology - importance

A

ability to visually determine which variations of DNA will cause expression of certain traits

42
Q

single nucleotide polymorphism

A

single base variations of DNA

42
Q

single nucleotide polymorphism

A

single base variations of DNA

43
Q

single nucleotide polymorphism - frequency

A

every 800 bases

43
Q

single nucleotide polymorphism - frequency

A

every 800 bases

44
Q

genome wide association studies

A

investigate genome as a whole, large patient samples, large control (normal) samples

44
Q

genome wide association studies

A

investigate genome as a whole, large patient samples, large control (normal) samples

45
Q

uses of microarrays [3]

A

[1] detect DNA variations
[2] detect changes in relative RNA quantity (degree of expression)
[3] detect changes in DNA methylation (degree of expression)

45
Q

uses of microarrays [3]

A

[1] detect DNA variations
[2] detect changes in relative RNA quantity (degree of expression)
[3] detect changes in DNA methylation (degree of expression)

46
Q

new generation sequencing

A

rapid sequencing technology making use of clonal amplification

46
Q

new generation sequencing

A

rapid sequencing technology making use of clonal amplification

47
Q

central to proteint testing

A

antibodies

47
Q

central to proteint testing

A

antibodies

48
Q

western blot

A

electrophoresis involving proteins

48
Q

western blot

A

electrophoresis involving proteins

49
Q

immunoassay

A

[1] antigens targeted by being attached to labeled antibodies
[2] competitive binding assay in which the binding protein being used is an antibody

49
Q

immunoassay

A

[1] antigens targeted by being attached to labeled antibodies
[2] competitive binding assay in which the binding protein being used is an antibody

50
Q

ELISA - meaning

A

enzyme-linked immunosorbent assay

50
Q

ELISA - meaning

A

enzyme-linked immunosorbent assay

51
Q

Interferon-gamma release assay - purpose

A

used to diagnose active/asymptomatic tuberculosis

51
Q

Interferon-gamma release assay - purpose

A

used to diagnose active/asymptomatic tuberculosis

52
Q

Interferon-gamma release assay - principle

A

immunocompetent T cells can be induced to express interferon-gamma only during active infection –> detection of this interferon gamma indicates infection

52
Q

Interferon-gamma release assay - principle

A

immunocompetent T cells can be induced to express interferon-gamma only during active infection –> detection of this interferon gamma indicates infection

53
Q

immunohistochemistry

A

[1] technique used to localize the antigens or proteins in tissue sections by use of labeled antibodies that are visualized by a marker such as fluorescent dyes
[2] tissue samples stained with antibodies specific to target proteins

53
Q

immunohistochemistry

A

[1] technique used to localize the antigens or proteins in tissue sections by use of labeled antibodies that are visualized by a marker such as fluorescent dyes
[2] tissue samples stained with antibodies specific to target proteins

54
Q

flourescent microscopy

A

technique using fluorescent antibodies as markers

54
Q

flourescent microscopy

A

technique using fluorescent antibodies as markers

55
Q

most of the differences in health conditions are associated with _____

A

variations in genomic structure and expression

55
Q

most of the differences in health conditions are associated with _____

A

variations in genomic structure and expression

[OS 201] 3rd Exam (10 decks)

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