technical methods- definitions

Question 1

basic principle of technical methods

Answer 1

choose the simplest, most direct method

Question 2

PCR- what does it do

Answer 2

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

Question 3

PCR uses

Answer 3

• 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

Question 4

qPCR- what does it do

Answer 4

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

Question 5

qPCR uses

Answer 5

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

Question 6

RNA seq- what does it do

Answer 6

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

Question 7

RNA seq uses

Answer 7

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

Question 8

DNA sequencing- what does it do

Answer 8

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

Question 9

DNA sequencing uses

Answer 9

mutation detections, pathogen identification, gene identification

Question 10

genome sequencing- what does it do

Answer 10

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

Question 11

genome sequencing uses

Answer 11

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

Question 12

GWAS - what does it do

Answer 12

use of single nucleotide polymorphisms to link genetic variation to disease

Question 13

GWAS uses

Answer 13

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

Question 14

trio analysis - what does it do

Answer 14

identification of disease- causing spontaneous mutations

Question 15

trio- analysis- uses

Answer 15

identification of the cause of rare genetic disease

Question 16

knock out- what does it do

Answer 16

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

Question 17

knock out uses

Answer 17

assessment of gene function- creating disease models

Question 18

CRISPR- what does it do

Answer 18

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

Question 19

CRISPR uses

Answer 19

creating knockouts. therapeutic repair of mutant genomes

Question 20

western blot what does it do

Answer 20

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

Question 21

western blot uses

Answer 21

determine changes in protein live. Disease/ tissue specific expression

Question 22

genetic screen- what does it do

Answer 22

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

Question 23

genetic screen uses

Answer 23

identification of genes important for a particular physiological or developmental process

Question 24

ELISA- what does it do

Answer 24

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

Question 25

ELISA- uses

Answer 25

determine changes i protein level

-detection of viral protein, antibody responses and of secreted molecules

Question 26

flow cytometry -what does it do

Answer 26

use of specific antibodies or fluorescent tags to detect proteins in intact cells

Question 27

flow cytometry uses

Answer 27

identification and quantification of different cell types e.g. separation of cell types- sorting

Question 28

immuno-fluorescence - what does it do

Answer 28

use of specific antibodies or fluorescent tags to detect proteins in intact cells using microscopy

Question 29

immunofluorescence uses

Answer 29

visual determination of which cells express specific proteins. sub-cellular localisation of proteins.