3.5 Genetic modification and biotechnology Flashcards

1
Q

What does PCR stand for?

A

Polymerase chain reaction

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2
Q

What is PCR?

A

An artificial method of replicating DNA in lab conditions

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3
Q

What is the PCR technique used to do?

A

Amplify large quantities of a specific sequence of DNA from an initial minute sample

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4
Q

What does each cycle of PCR do to the amount of DNA?

A

Double it

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5
Q

Where does PCR occur?

A

In a thermal cycler

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6
Q

What does PCR use to control the replication process?

A

Variations in temperature

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7
Q

What are the three stages of PCR?

A

Denaturation
Annealing
Elongation

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8
Q

What happens during the denaturation stage of PCR?

A

DNA sample is heated to separate it into two single strands

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9
Q

What is the temperature for the denaturation stage of PCR?

A

95C for 1 min

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10
Q

What happens during the annealing stage of PCR?

A

DNA primers attach to the 3’ ends of the target sequence

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11
Q

What is the temperature during the annealing stage of PCR?

A

55C for 1 min

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12
Q

What happens during the elongation stage of PCR?

A

A heat tolerant DNA polymerase binds to the primer and copies the strand

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13
Q

What is the temperature used during the elongation stage of PCR?

A

72C for 2 min

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14
Q

After PCR and once large quantities of DNA have been created what happens?

A

Other lab techniques are used to isolate and manipulate the sequences

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15
Q

What is gel electrophoresis?

A

A lab technique used to separate and isolate proteins or DNA fragments based on size

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16
Q

What happens in gel electrophoresis?

A

Samples are placed in a block of gel and an electric current is applied which causes the samples to move through the gel

17
Q

Why will smaller samples move faster through the gel in gel electrophoresis?

A

As they are less impeded by the gel matrix

18
Q

Why do samples of different sizes separate?

A

As they travel at different speeds

19
Q

How may DNA be cut into fragments?

A

Using restriction endonuclease

20
Q

What will different DNA samples generate?

A

Different fragment lengths

21
Q

In DNA separation, why do fragments separate?

A

Because DNA is negatively charged due to the presence of a phosphate group on each nucleotide

22
Q

In DNA separation, where are DNA samples placed?

A

Into an agarose gel

23
Q

In DNA separation, how is fragment size calculated?

A

By comparing against known industry standards

24
Q

In DNA separation how can specific sequences be indentified?

A

By incorporating a complementary radiolabelled hybridisation probe, transferring the separated sequences to a membrane and then visualising via autoradiography

25
In protein separation, what must proteins first be treated with?
An anionic detergent
26
In protein separation,why must proteins be treated with an anionic detergent?
To linearise and impart a uniform negative charge
27
In protein separation, where are protein samples placed?
In polyacrylamide gel
28
In protein separation, what are sizes compared against?
Known industry standards
29
In protein separation, where are separated proteins transferred to?
A membrane
30
In protein separation, how are target proteins identified?
By staining with specific monoclonal antibodies
31
How can proteins affect size?
They may be folded into a variety of shapes
32
How do proteins have no clear charge?
They have positive and negative regions
33