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3. PCR Page 376 – 377
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PCR Polymerase Chain Reaction PCR – in vitro
The more DNA available, the easier it is to work with.
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Kary Mullis Developed the PCR process in 1986 Nobel Laureate, 1993
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Polymerase Chain Reaction
Uses purified DNA polymerase Minimum requirements for DNA polymerase:
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Three steps for a cycle DNA strand denaturation (95°C)
Primer annealing (50°C - 65°C) DNA strand synthesis (72°C)
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PCR
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Problem Where do you find enzymes that don’t break down at 95°C? Thermus aquaticus Isolated Taq polymerase from these bacteria.
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PCR After 30 cycles, 230 (more than a billion) copies of DNA can be produced. 30 cycles of PCR can take anywhere from 1 – 2 hours to complete.
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PCR Applications Genetic Screening Forensic Analysis
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Video PCR Animation
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