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

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Flashcards in Trans - Utility of Molecular Tools in Health Care and Biological Research Deck (83)
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1

discovered DNA double helix

James Watson, Francis Crick

2

BRCA 1 & 2

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

3

physical manifestation of genetic variation

anatomic variation

4

biochemical manifestation of genetic variation

physiologic

5

main genomic variations [3]

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

6

epigenetic mechanism of genetic variation

[1] methylation of DNA
[2] histone packing

7

epigenetic mechanism affected by: [5]

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

8

methods of nucleic acid extraction

[1] organic-based extraction
[2] silica-based extraction

9

methods of nucleic acid amplification

[1] polymerase chain reaction
[2] cloning

10

post-amplification methods [4]

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

11

when will you use organic methods of nucleic acid extraction?

when you want a greater quantity of nucleic acid

12

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

when you want greater purity of the sample

13

principle of nucleic acid extraction by organic solvent

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

14

2 main organic solvents used in nucleic acid extraction

[1] chloroform
[2] alcohol

15

requirements of PCR [4]

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

16

PCR - temperature to activate DNA polymerase

94-96 C

17

PCR - denaturation step temperature

94-96C

18

PCR - annealing step - temperature and purpose

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

19

PCR - extension step - temperature and purpose

72 C to allow elongation

20

PCR - final extension step - temperature and purpose

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

21

PCR - final hold - temperature and purpose

4-15 C for short-term storage

22

DNA polymerase used in PCR? why?

Taq polymerase - heat resistant polymerase from Thermus aquaticus

23

electrophoresis - principle

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

24

electrophoresis - purpose

to obtain a certain DNA segment of interest

25

electrophoresis - direction of flow? why?

towards the + terminal, because DNA is negatively charged

26

electrophoresis - rate of flow is affected by what factors?

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

27

LAMP - meaning

Loop Mediated Isothermal Amplification of DNA

28

LAMP - principle

PCR conducted at only one temperature

28

LAMP - principle

PCR conducted at only one temperature

29

LAMP - microorganism involved

bacillus stereos thermophiles (BST)