technical methods- definitions Flashcards

1
Q

basic principle of technical methods

A

choose the simplest, most direct method

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2
Q

PCR- what does it do

A

amplifies a specific DNA sequence determine day the experimenter. Specific and very sensitive

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3
Q

PCR uses

A
  • Detection of viral and bacterial DNA in tissue samples
  • Determining the size of a sequence of DNA between two ‘primers’
  • production of DNA for making genetic tools (e.g. expression vectors) or DNA sequencing
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4
Q

qPCR- what does it do

A

quantifies a specific DNA (determined by the experimenter) sequence in a sample

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5
Q

qPCR uses

A
  • quantifying viral load
  • comparing amounts of DNA
  • comparing amounts of specific mRNA (following conversion to DNA by RT)
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6
Q

RNA seq- what does it do

A

quantifies all RNA species in a sample- converts all RNA to DNA and then uses parallel sequencing to identify each molecule

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7
Q

RNA seq uses

A

comparative gene expression. Finding out which genes are regulated in a particular disease or in response to a specific stimulus. An un-bias method

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8
Q

DNA sequencing- what does it do

A

determines the base sequence of a specific stretch of DNA (sound 750 bases)

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9
Q

DNA sequencing uses

A

mutation detections, pathogen identification, gene identification

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10
Q

genome sequencing- what does it do

A

massive parallel DNA sequencing to establish the base sequence of the entire genome

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11
Q

genome sequencing uses

A

gaining all at a on genetic variation. Identification of spontaneous mutation

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12
Q

GWAS - what does it do

A

use of single nucleotide polymorphisms to link genetic variation to disease

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13
Q

GWAS uses

A

understanding pathways involved in complex disease. assessment of individuals disease risk

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14
Q

trio analysis - what does it do

A

identification of disease- causing spontaneous mutations

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15
Q

trio- analysis- uses

A

identification of the cause of rare genetic disease

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16
Q

knock out- what does it do

A

commonly in mice or model organism/ cell lines- disruption and inactivation of a specific target gene

17
Q

knock out uses

A

assessment of gene function- creating disease models

18
Q

CRISPR- what does it do

A

genome editing. allows user targeted disruption or alteration of mammalian genome

19
Q

CRISPR uses

A

creating knockouts. therapeutic repair of mutant genomes

20
Q

western blot what does it do

A

use of specific antibody to detect and quantify a particular protein in a sample

21
Q

western blot uses

A

determine changes in protein live. Disease/ tissue specific expression

22
Q

genetic screen- what does it do

A

random low level mutation of large numbers of a model organic,s

23
Q

genetic screen uses

A

identification of genes important for a particular physiological or developmental process

24
Q

ELISA- what does it do

A

use of specific antibodies or antigen to detect and quantify a particular protein or molecule in a sample

25
ELISA- uses
determine changes i protein level | -detection of viral protein, antibody responses and of secreted molecules
26
flow cytometry -what does it do
use of specific antibodies or fluorescent tags to detect proteins in intact cells
27
flow cytometry uses
identification and quantification of different cell types e.g. separation of cell types- sorting
28
immuno-fluorescence - what does it do
use of specific antibodies or fluorescent tags to detect proteins in intact cells using microscopy
29
immunofluorescence uses
visual determination of which cells express specific proteins. sub-cellular localisation of proteins.