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GDAN year 2 G&D with some aging > Basic genetics in relation to DNA gene reg, gene expression and polymorphisms > Flashcards

Basic genetics in relation to DNA gene reg, gene expression and polymorphisms Flashcards

1
Q

what does DNA finger printing rely on

A

variabel number of tandem repeats

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2
Q

myoglobin gene

A

segments in introns with no apparent function

- vary in size/composition between individuals

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3
Q

enzyme which cuts DNA

A

restriction enzyme

cuts at specific sequences

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4
Q

electrophoresis

A

run DNA fragments through gel
pass electric current accross
smaller the molecule faster it will run

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5
Q

what does gel electrophoresis separate fragments by

A

size

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6
Q

starting material for PCR

A

DNA

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7
Q

how do you reverse transcripbe to amplify

A

use RNA to make cDNA

polymerase chain reaction to make copies

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8
Q

polymerise chain reaction steps

A

open DNA by heating to 94 degrees, uncoils
add primers(forward and reverse), join at every end of DNA and corresponds
reduce temp to 54
extension occurs at 72 where new DNA is built

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9
Q

what is hybridisation

A

DNA sticks to DNA and RNA

RNA sticks to DNA and RNA

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10
Q

what can you do after amplification

A

run sequences on gel

show gene expression being associated with phenotype o the cell

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11
Q

problems with end point PCR

A
  • poor precision
  • lower sensitivity
  • short dynamic range
  • low resolution
  • non automated
  • size based discrimination only
  • results not expressed as numbers
  • ethidium bromide staining is not very quantitive (likes to stick to nuclear material)
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12
Q

what does real time PCR detect

A

accumulation of amplicon during reaction

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13
Q

what is the optimal point for analysing data in real time RCR

A

exponential phase

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14
Q

why can you not take data at the plateau in real time PCR

A

different amounts of DNA in the starting material

- data would not truly represent starting materail

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15
Q

what are the methods for real time PR

A

Taq man probe

flurorences

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16
Q

Taq man probe method

A

has a quenching and reporter dye

  • Taq man probe and primer binds to target DNA
  • Taq polymerase can now create complementary strand
  • Reported dye released from the extending double stranded DNA
  • reporter dye now distant to quencher so it will fluoresce and give a signal
  • machine analyses this
17
Q

what does light relate to in taq man probe method

A

amount of light relates to amount of DNA in sample

18
Q

2nd method for real time PCR

A
  • cyber green attatches to any double stranded DNA
  • When the DNA is denatured the green floats free
  • can then pick up the fluorescent activity by the machine
19
Q

advantages of real time PCD

A

traditional measure at end point and real time collects date in exponential growth phase
increase in reported fluorescence directly proportional to number of amplicons generated
increased dynamic range of detection
no post PCR processing

20
Q

transcriptomics

A

genome wide RNA transcript expression levesl

21
Q

in situ hydribdication

A

localisation technique for the detection of a gene product (RNA) in tissues

22
Q

what can in situ hybridisation be used for

A

investigate cell type and tissue specific gene expression

23
Q

genomics includes..

A

recombinant DNA
DNA sequencing methods
bioinformatics
- used to sequence, assemble and analyse the function/strucute of the genme

24
Q

proteomics

A

large scale study of proteis

25
Q

metabolomics

A

comprehensive characterisation of small molecule metabolites in biological systems

26
Q

what is a nucleotide polymorphsm

A

single nucleotide change in genome sequence

some no effect, some lead to disease

27
Q

genome wide associated studies

A

rapid scanning of complete sets of DNA of many people to find genetic variations associated with a particular disease

28
Q

Taq man probe method

A

has a quenching and reporter dye

  • Taq man probe and primer binds to target DNA
  • Taq polymerase can now create complementary strand
  • Reported dye released from the extending double stranded DNA
  • reporter dye now distant to quencher so it will fluoresce and give a signal
  • machine analyses this
29
Q

what does light relate to in taq man probe method

A

amount of light relates to amount of DNA in sample

30
Q

2nd method for real time PCR

A
  • cyber green attatches to any double stranded DNA
  • When the DNA is denatured the green floats free
  • can then pick up the fluorescent activity by the machine
31
Q

advantages of real time PCD

A

traditional measure at end point and real time collects date in exponential growth phase
increase in reported fluorescence directly proportional to number of amplicons generated
increased dynamic range of detection
no post PCR processing

32
Q

transcriptomics

A

genome wide RNA transcript expression levesl

33
Q

in situ hydribdication

A

localisation technique for the detection of a gene product (RNA) in tissues

34
Q

what can in situ hybridisation be used for

A

investigate cell type and tissue specific gene expression

35
Q

genomics includes..

A

recombinant DNA
DNA sequencing methods
bioinformatics
- used to sequence, assemble and analyse the function/strucute of the genme

36
Q

proteomics

A

large scale study of proteis

37
Q

metabolomics

A

comprehensive characterisation of small molecule metabolites in biological systems

38
Q

what is a nucleotide polymorphsm

A

single nucleotide change in genome sequence

some no effect, some lead to disease

39
Q

genome wide associated studies

A

rapid scanning of complete sets of DNA of many people to find genetic variations associated with a particular disease

GDAN year 2 G&D with some aging (27 decks)

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