In vitro cloning - polymerase chain reaction.

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What does PCR stand for?
The polymerase chain reaction.
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Where is DNA polymerase obtained from?
Bacteria that live in hot springs.
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What are primers?
Short sequences of nucleotides that have a set of bases complimentary to those at one end of the two DNA fragments.
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What is a thermorecycler?
A computer controlled machine that varies temperature precisely over a period of time.
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What are the three stages of PCR?
1. Separation of the DNA strand. 2. Addition (annealing) of primers. 3. Synthesis of DNA.
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What happens in the separation of the DNA strand?
DNA fragments, primers and DNA polymerase are placed in a vessel in a thermocycler. Temp is increased to 95 degrees, causing the DNA fragments to separate.
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What happens in annealing of the primers?
Mixture is cooled to 55 degrees causing primers to join to their complimentary bases. The primers provide starting sequences for DNA polymerase to begin copying.
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What happens in synthesis of DNA?
Temp is increased to 72 degrees. This is optimum temperature for DNA polymerase to add complimentary nucleotides along each of the DNA strands. It begins at the primer on both strands and adds the nucleotides in sequence until it reaches the end.
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Give two strengths of in vitro fertilisation?
1. It is rapid which is useful when analysing the scene of a crime. 2. It does not require living cells all that it requires is a base sequence of DNA.
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Give 3 strengths of in vivo fertilisation?
1. Useful when we want to introduce a gene into another organism. 2. No risk of contamination, same restriction endonuclease is used. 3. It is very precise as it cuts out specific genes.
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Card 2

Front

Where is DNA polymerase obtained from?

Back

Bacteria that live in hot springs.

Card 3

Front

What are primers?

Back

Preview of the front of card 3

Card 4

Front

What is a thermorecycler?

Back

Preview of the front of card 4

Card 5

Front

What are the three stages of PCR?

Back

Preview of the front of card 5
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