Gene technology Flashcards

1
Q

How can DNA fragments be made?

A

Conversion of mRNA to DNA using reverse transcriptase (cDNA)
Cut out DNA section using restriction endonucleases
Create a gene in a gene machine

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2
Q

What does in vitro mean?

A

Outside a living organism

In glassware

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3
Q

What does in vivo mean?

A

Carried out inside a living organism

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4
Q

For what is PCR an abbreviation?

A

Polymerase chain reaction

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5
Q

List the items needed to perform PCR

A
Thermocycler
Primers
DNA Nucleotides
DNA polymerase
Section of DNA to copy
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6
Q

In PCR the thermocycler initially heats the contents to 95 oC. What happens at this temperature?

A

DNA double helix separates into single strands

Hydrogen bonds between complementary base pairs are broken

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7
Q

What is a primer?

A

A short piece of DNA (15-20 bases long) with complementary base sequence to the ends of the DNA fragment you wish to copy

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8
Q

At what temperature do primers bind to the DNA fragments?

A

55oC

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9
Q

What is taq DNA polymerase?

A

DNA polymerase that is not denatured at high temperatures.

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10
Q

Where is taq DNA obtained from?

A

Thermophilic bacteria which live in hot springs

eg Thermus aquaticus

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11
Q

Why is a thermocycler used in PCR?

A

A computer controlled machine which allows temperatures to be controlled and changed quickly and accurately
Cycle take about 2 minutes

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12
Q

What is the function of the DNA polymerase in a PCR machine

A

Catalyses the formation of phosphodiester bonds between nucleotides

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13
Q

What is the advantage of using in vitro cloning?

A

Quick

No living cells needed

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14
Q

Give the sequence of temperatures used in PCR and state what happens at each temperature

A

95oC - breaks hydrogen bonds between DNA strands
55oC - primers anneal to DNA strands
72oC - DNA polymerase catalyses the formation of phosphodiester bonds between nucleotides

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