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6.1.3 Manipulating Genomes > DNA Profiling > Flashcards

DNA Profiling Flashcards

1
Q

What is a genome?

A

-all the genes possessed by an individual or an organism
-99% the same across humans

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2
Q

What are exons?

A

-code for proteins

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3
Q

What are introns?

A

-non coding sections of DNA
-gene expression control

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4
Q

What is satellite DNA?

A

-repeated sequences in introns, centromeres + telomeres

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5
Q

What are the 2 types of satellite DNA?

A
  1. Mini
  2. Micro
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6
Q

What is minisatellite DNA?

A

-VNTR’s
-20-50 base paris repeated 50-100 times

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7
Q

What is microsatellite DNA?

A

-STR’s
-2-4 base pairs repeated 5-15 times

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8
Q

What is a telomere?

A

-repeated at the end
-where satellites are found
-diff people have diff no.of repeats = satellite patterns

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9
Q

What does DNA profiling help do?

A

-helps identify an individual + determine familial relationships

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10
Q

What are the 7 steps of DNA profiling?

A
  1. Extract DNA
  2. PCR
  3. Restrictive endonucleases
  4. Electrophoresis
  5. Southern blotting
  6. Hybridisation
  7. Seeing evidence
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11
Q

What does PCR do?

A

-amplify DNA sample

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12
Q

What is PCR?

A

-artificial DNA replication
-to amplify DNA fragment for further processing

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13
Q

What happens during restrictive endonucleases?

A

-cut large fragments into satellites
-they have active complementary site to the base
-in introns (satellites intact)

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14
Q

What happens during electrophoresis?

A

-agar jelly emerged in alkaline to break H bonds
-strands placed at negative end

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15
Q

Why are the strands placed at negative pole during electrophoresis?

A

-DNA has a phosphate group which = negative
-repelled from - and travels to +

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16
Q

What happens during southern blotting?

A

-blot DNA onto nylon sheet
-transfers pattern

17
Q

What happens during hybridisation?

A

-add dye
-heat + UV light to fix pattern in place
=evidence to see

18
Q

What are probes?

A

-fragments of DNA or RNA complementary to satellites that were selected
-can be radioactive or florescent

19
Q

What are primers?

A

-short DNA sequences that anneal to DNA section that is wanted

20
Q

What are the 3 stages of PCR?

A
  1. Denaturation
  2. Annealing of primers
  3. Extending/synthesis
21
Q

What happens during denaturation?

A

-increase KE = vibration
-bases separation from their complementary bases
-breaks H bonds

22
Q

What temperature does denaturation occur at?

A

95 c

23
Q

What happens during the annealment of primers?

A

-design primers to recognise target DNA
-to be amplified

24
Q

What temperature does annealing occur at?

A

55 c

25
Q

What happens during extending/synthesis?

A

-free nucleotides air up with exposed bases
-tag polymerase joins up backbone in 5’ to 3’ direction
-forming phosphodiester bonds

26
Q

What temperature does extending/synthesis occur at?

A

72 c

6.1.3 Manipulating Genomes (6 decks)

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