14 pcr

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14 pcr

  1. 1. pp 382 - 383
  2. 2. Kary Mullis Developed the PCR process in 1986 Nobel Laureate, 1993
  3. 3. Polymerase Chain Reaction PCR – a synthetic method (no living cells) to produce millions of copies of DNA  use enzymes in a tube - in vitro The more DNA available, the easier it is to work with. Minimum requirements for DNA polymerase: 1. 2. 3. template strand primers dNTPs
  4. 4. Three Steps for each PCR Cycle 1. DNA strand denaturation (95°C) separate double stranded DNA  each strand becomes template strand  1. Primer annealing  1. bind short DNA pieces to template strands DNA strand synthesis  (50°C - 65°C) produce new DNA strands (72°C)
  5. 5. Problem Where do you find enzymes that don’t break down at 95°C? Thermus aquaticus bacterium live in hot springs and their enzymes are designed to withstand extreme temperatures. Isolated Taq polymerase from these bacteria.
  6. 6. 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.
  7. 7. PCR
  8. 8. PCR Applications Genetic Screening Early detection for certain diseases. Forensic Analysis Small amounts of DNA collected to identify individuals.
  9. 9. PCR Animation

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