lab research

Question 1

why is laboratory-based biomedical research important?

Answer 1

• not all questions can be answered using humans

Question 2

5 things that laboratory-based biomedical research includes?

Answer 2

1.exploring new phenomena
2.understanding diseases mechanisms
3.testing new hypotheses
4.screening for inhibitors
5.identifying targets/candidates

Question 3

lab research vs field research

Answer 3

-controlled- variables can be —well controlled
reproducibility
-clear conclusions
-artificial/ non physiological settings
-may not translate well to humans
-expensive

Question 4

8 steps of process of lab research

Answer 4

1.defining research Q
2.defines aims of project
3.design and plan experiments
4.order reagents and equipment
5.tool or method development
6.tool or method development
7.conduct experiments
8.data analysis
9.conclusions

Question 5

what 2 process used for DNA

Answer 5

PCR, genome sequencing

Question 6

what 2 process is used for

Answer 6

RT-PCR , RNAsea

Question 7

what 3 process used for protein?

Answer 7

1.PAGE
2.proteotamics
3.immunblotting

Question 8

DNA extraction

Answer 8

methods involve disruption of cells using lysis regents and subsequent treatment with different chemicals remove other biomolecules

Question 9

common method in DNA extraction

Answer 9

organic extraction using phenomenal and chloroform, both mixtures denature and precipitate out portions and treatments removed the cellular RNA

Question 10

2 Specialised protocols used for extraction of high quality DNA from clinical samples

Answer 10

1.blood
2.feaces

Question 11

Polymerase chain reaction(PCR)

Answer 11

new development and modification to increase accuracy and efficiency an enzyme

Question 12

6 PCR features

Answer 12

1.cloning
2.disease diagnosis
3.detection of infectious agents
4.forensics
5.evolutionary biology
6.Archeology-ancient DNA anaylsis

Question 13

Genome sequencing

Answer 13

DNA sequence of the genome of an organism

Question 14

4 basics of whole genome(shotgun) sequencing

Answer 14

-high quality genomic DNA
-sheared to small fragments
-tagged with adapted molecules and amplified
sequenced in a sequencing machine such as illuminating

Question 15

6 applications of illumination sequencing

Answer 15

1.disease diagnosis/predication in clinical medicine
2.prenatal, newborn screening
3.infectious disease epidemiology
4.bacterial, viral evolution studies
5.human population history studies
6.personalised medicine

Question 16

RNA(reverse transcriptionPCR)

Answer 16

synthesis of complementary DNA from RNA

Question 17

What does reverse transcriptase occur in?

Answer 17

RNA viruses(retroviruses)-mediates conversion from RNA to DNA

Question 18

what happens in RT-PCR?

Answer 18

reverse transcription is coupled with PCR

Question 19

3 Applications of RT-PCR

Answer 19

1.generation of cDNA for cloning and cDNA libraries
2.recombinant protein expression
3.key to several global transcriptomic methodologies e.g. RNA seq

Question 20

3 things RNAseq does?

Answer 20

1.sequencing all transcripts in a cell
2.global view/snapshot of which genes are on/off at a time
3.quantitate gene expression

Question 21

is the PCR method quantitative or qualitative?

Answer 21

quanitative-is more sensitive and specific

Question 22

5 applications of PCR

Answer 22

1.disease diagnosis-quantitate specific transcripts
2.measure microbial load
3.gene expression studies
4.SNP analysis
5.genotyping

Question 23

Polacrylamide gel electrophoresis(PAGE)

Answer 23

basic method for analysing cellular protein

Question 24

4 steps of process in SDS-PAGE

Answer 24

1.denaturing proteins using detergent sodium dodecyl sulphate(SDS)
2.elimates structure and charge of protiens
3.separation based only on molecular weight of polypeptides
4.widely used to separate proteins and visualise protein profiles

Question 25

3 things western-blotting(immunoblotting) does?

Answer 25

1.protiens on membranes are detected using specific antibodies
2.high sensitivity detects low amount of proteins
3.detect specific proteins and quantitate proteins

Question 26

what does ELISA(enzyme-linked immunosorbent assay) measure?

Answer 26

antibodies, or specific protiens from biological samples

Question 27

3 applications of ELISA

Answer 27

1.HIV detection
2.measuring cytokines
3.pregnancy test

Question 28

in Proteomics what is proteome?

Answer 28

set of proteins produced by a cell, system or organism under a set of conditions

Question 29

what is proteome used to study?

Answer 29

protein production
stability
control
protein modifications
protein interactions
protein movement across sub cellular compartments

Question 30

in vitro cellular models

Answer 30

-cell lines or primary cells

Question 31

in vitro cellular models, what are primary cells derived from

Answer 31

tissues

Question 32

in vitro cellular models, what are cell lines?

Answer 32

cells are transformed to allow continuous propagation

Question 33

3 things in vitro cellular modals do?

Answer 33

1.genetically manipulate
2.known out genes
3.visualise proteins(florescent labelled)

Question 34

what does in vitro cellular assays study?

Answer 34

human responses

Question 35

2 functions of in vitro cellular assays ?

Answer 35

1.measurement of cellular morphology
2.cell proliferation

Question 36

3 advantages of animal modals

Answer 36

-inexpensive and easy to handle
-gene knockout strains available
-immunological regents available

Question 37

disadvantages of in vitro assays

Answer 37

-findings may not always be relevant to tissue/whole organism

Question 38

disadvantages of in-vivo assays

Answer 38

animals respond differently to infection -diiferent immune and cellular responses

Question 39

disadvantages of lab research

Answer 39

funding limitations impact use of techniques
some techniques need extensive training
when applying multiple methods to answer the same question they may not give the same result