Lecture 21 Flashcards

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1
Q

Who invented PCR?

A

Kary Mullis in 1970s

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2
Q

PCR is based on 2 key concepts, which ones?

A
  1. When DNA is expose to high temperatures, the hydrogen bonds that hold the two strands together will break, giving 2 single strands of DNA.
  2. Extremophiles live and can carryout cellular processes at very high temperatures.
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3
Q

What is DNA polymerase?

A

the enzyme responsible for copying the sequence starting at the obligonucleotide primer from the single DNA strand

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4
Q

What is PCR?

A

it is an in vitro technique for the amplification of a region of DNA which lies between 2 regions of known sequence

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5
Q

At which cycle does the target DNA appears?

A

at the third cycle

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6
Q

PCR grows…

A

exponentially

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7
Q

What do you need to carry out PCR?

A
  • template DNA
  • free nucleotides
  • heat stable polymerase
  • thermocycler
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8
Q

What are the 3 steps of PCR?

A
  1. Denaturation: the DNA samples are heated at 95°C and denatured
  2. Annealing: the temperature is cooled (45-54 °C) and the primers anneal to the specific region
  3. Extension: Taq polymerase extends the DNA (72°C) thus giving new copies of the DNA

www.youtube.com/watch?v=iQsu3Kz9NYo

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9
Q

PCR cannot amplify large amounts of DNA, only used on smaller specific regions such as genes (10,000 bp)

A

true

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