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214.102 Applied Sciences for Health Professionals > Module 2 - Genetic techniques > Flashcards

Module 2 - Genetic techniques Flashcards

1
Q

How can knowing about the DNA sequence/structure help us?

A

Used for

  • producing drugs, vaccines
  • identifying if you have a genetic disease
  • helping to cure genetic disease
  • forensics
  • genetic screening and DNA testing
  • genetic counselling
  • Research
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2
Q

PCR

A

Polymerase Chain Reaction

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3
Q

PCR Allows specific region of DNA to be —————- exponentially from minute quantity templates

A

Amplified

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4
Q

PCR Offers alternative approach to —- ——- for the production of many copies of identical sequence of DNA

A

Gene cloning

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5
Q

IN PCR starting material may be genomic —, —, ————– ————- DNA, ——– DNA or ————- —————

A 
DNA
RNA
Archival specimen
Cloned
Forensic samples
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6
Q

enzymes which can make DNA from dNTPs provided a template or primer is available to start the process

A

Polymerase

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7
Q

First step in PCR

A

denaturation of double strand of DNA

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8
Q

Second step in PCR

A

annealing of primers

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9
Q

Third step in PCR

A

elongation of strands

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10
Q

cDNA

A

Copy of DNA

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11
Q

Denaturation of dsDNA - typically temp used is ————— for human genomic DNA.

A

93-95 degrees C

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12
Q

Annealing - temp ————-, depending on melting temperature of the expected duplex.

A

40-70 degrees C

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13
Q

DNA Synthesis - generally ———–: use the optimum temperature for the polymerase being used.

A

70-75 degrees C

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14
Q

Product size = Length of ————–/—————/left ————– + Length of ———–/———————–/right —————- + —————- to be amplified

A

Forward, upstream, primer
Reverse, downstream, primer
Region

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15
Q

PCR:
DNA is ————–
————Attach to each strand
A new DNA strand is ——————– ————- primers on each template strand

A

denatured
Primers
Synthesised
Behind

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16
Q

Continued rounds of —————–swiftly produce large numbers of identical fragments

A

Amplification

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17
Q

Applications of PCR: D———— and S————- of genetic diseases and cancer

A

Diagnosis; Screening

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18
Q

Applications of PCR: Rapid detection of slowly growing ————————-

A

Microorganisms

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19
Q

Applications of PCR: — typing in T———————–

A

HLA; transplantation

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20
Q

Applications of PCR: Analysis of DNA in A———– M————-

A

Archival material

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21
Q

Applications of PCR: DNA F——————— in forensic science

A

Fingerprinting

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22
Q

Applications of PCR: Preparation of nucleic acid P————-

A

Probes

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23
Q

Applications of PCR: ———— screening, mapping and subcloning

A

Clone

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24
Q

detection of mRNA expression levels

A

RT-PCR

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25
Q

RT-PCR traditionally involves —– steps

A

two

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26
Q

Step 1. the —— reaction and ———– amplification. RNA is first reverse transcribed into ——— using a reverse ———————

A

RT; PCR; cDNA; transcriptase

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27
Q

Step 2. the resulting —– is used as templates for subsequent ————- using ———— specific for one or more genes.

A

cDNA; PCR amplification; primers

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28
Q

Genetic Techniques 1

A

Diagnostic PCR- template DNA

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29
Q

Genetic Techniques 2

A

RT-PCR – template RNA, can be diagnostic

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30
Q

Genetic Techniques 3

A

DNA sequencing

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31
Q

Process of manipulating genes, usually outside the organisms normal reproductive process.

A

Genetic engineering

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32
Q

Genetic engineering Usually involves ———–, ——————– and —————— of DNA into cells or model organisms usually to express a —————

A

isolation; manipulation; reintroduction; protein

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33
Q

Genetic Engineering Techniques: 3 examples

A

Recombinant DNA,
cloning
gene therapy

34
Q

What two things are genetic engineering techniques used for?

A

production of drugs, vaccines, treatment strategies

research e.g. investigating mechanisms of disease

35
Q

Recombinant DNA technology: Basic procedure 1: DNA ———– from cells or tissues

A

purified

36
Q

Recombinant DNA technology: Basic procedure 2: ———— —————- used to generate specific DNA fragments, recognise and cut DNA at specific sequence

A

Restriction enzymes

37
Q

Recombinant DNA technology: Basic procedure 3: Fragments join to other DNA molecules which act as ————- (carrier molecules), —————- + DNA fragment= ————— DNA molecule

A

vectors; vector; recombinant

38
Q

Recombinant DNA technology: Basic procedure 4: Recombinant DNA molecule transferred to ——- ———

A

host cell

39
Q

Recombinant DNA technology: Basic procedure 5: Within host cell recombinant ————— producing dozens of identical copies

A

replicates

40
Q

Recombinant DNA -As host replicates, recombinant molecule passed to all ————– creating population of cells each carrying cloned DNA sequence

A

progeny

41
Q

Recombinant DNA technology: Basic procedure 7: Cloned DNA can be recovered from host cells, ———– and —————

A

purified; analysed

42
Q

Recombinant DNA technology: Basic procedure 8: Cloned DNA in host cells is ——————, its mRNA —————-, gene product ————- and used or sold

A

transcribed; translated; isolated

43
Q

First genetically engineered drug =

A

human insulin (1982)

44
Q

First genetically engineered vaccine =

A

hepatitis B (1986)

45
Q

the creation of an exact genetic replica of a small segment of DNA, a cell or a whole organism

A

Reproductive cloning

46
Q

stem cells

A

Therapeutic cloning

47
Q

Cloning = —————- ————- cell + egg cell with ————— removed

A

Somatic body; Nucleus

48
Q

Nucleus of somatic body cell fused with denucelated egg cell

A

cloning

49
Q

reproductive cloning - —————- mother

A

surrogate

50
Q

Therapeutic cloning —> ———— ——————–

A

Tissue culture

51
Q

Dolly the sheep - Cloned in —– - Died in ————

A

1997; 2003

52
Q

a technique for correcting defective genes responsible for disease development

A

Gene Therapy

53
Q

In most cases of gene therapy, a “normal” gene is ————- into the genome to ————- an “abnormal” disease causing gene.

A

inserted; replace

54
Q

There are two major techniques in gene therapy

A

Gene Augmentation Therapy

Targeted Modulation of gene expression

55
Q

The introduction of DNA into a cell with a view to augmenting gene function, e.g. recessive diseases.

A

Gene Augmentation Therapy

56
Q

could also be used to deliver genes encoding proteins that would be responsible for stimulating the immune system to treat cancer or infections

A

Gene Augmentation Therapy

57
Q

Methods for turning off genes using for example ribozymes and anti-sense oligonucleotides, e.g., infectious disease and cancers (later lecture).

A

Targeted Modulation of gene expression

58
Q

A ———– needed to deliver the therapeutic gene to the patients target cells

A

vector

59
Q

Gene therapy: Most common vector is ——-

A

virus

60
Q

First human gene therapy experiment when?

A

1990

61
Q

Gene therapy is not really successful in humans due to the lack of efficient ———- ————- for gene delivery.

A

vector systems

62
Q

DNA delivered to cells can be ———– and in the case of cystic fibrosis restore —————

A

expressed; function

63
Q

Gene therapy: Shift in emphasis to ———- and ——– rather than ‘genetic diseases

A

Cancer; HIV

64
Q

DNA testing 1: collect a persons ————— ———

A

total DNA

65
Q

DNA testing 2: DNA cut into small pieces with ————– that are chemical scissors

A

enzymes

66
Q

DNA testing 3: DNA fragments put on ——

A

gel

67
Q

DNA testing 4: fragments separated according to ———– using an ———– ———–

A

size; electrical current

68
Q

DNA testing 5: Separated fragments transferred to ———— —————– and mixed with DNA ———– that picks up the regions of DNA interest

A

nylon membrane; probe

69
Q

Provides information and support to an individual or family about genetic disorders

A

Genetic Counselling

70
Q

Who uses Genetic Counselling?

A

A child is born with several abnormalities
There is a history in the family of a medical condition
A woman has a history of several miscarriages
Woman aged over 35 yrs and pregnant
Anyone who has uncertainties about genetic risks

71
Q

the examination of a person’s chromosomes, DNA or the biochemical product of a gene (protein)

A

Genetic Testing

72
Q

Tissues tested in genetic testing:

A

blood; skin; saliva; hair follicles; amniotic; CVS; placental

73
Q

CVS test done in pregnancy around —— weeks

A

11-12

74
Q

Amniocentesis done in pregnancy around ——– weeks

A

15-19

75
Q

CVS = ————– tissue withdrawn

A

chorionic

76
Q

Amneocentesis = ————- ————– withdrawn

A

amniotic fluid

77
Q

3 types of Genetic Tests Used

A

Cytogenic testing - examine chromosomes
DNA testing – DNA extracted from cells of a variety of body fluids or tissues
Proteins – usually tested for in blood

78
Q

Research: Genetic techniques used all the time to ——————- —————-

A

investigate diseases

79
Q

Research: Identify ———- and ————- —————-

A

therapeutic; diagnostic markers

80
Q

Research: Understand ———— ————- involved in development of diseases

A

molecular mechanisms

81
Q

Research: Identify genes that are ——————- and cause disease

A

abnormal

214.102 Applied Sciences for Health Professionals (14 decks)

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