PCR

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PCR

• Polymerase Chain Reaction

Revised 10-31-2006
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The Nobel Prize in Chemistry 1993

• Kary B. Mullis
• "for contributions to the developments of methods
  within DNA-based chemistry" "for his invention of
  the polymerase chain reaction (PCR) method

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Taq polymerase
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Thermus aquaticus
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Thermal Cycler
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Materials of PCR

• target DNA
• Taq DNA polymerase
• 2 Primers
• 20 nucleotides in length
• Forward and reverse
• the four DNTPS
• Adenine
• Thymine
• Cytosine
• Guanine
• cofactor MgCl2.

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Locate the Target Sequence

• Scientists determine which GENE they are
  interested in studying
• Locate Primers Upstream and Down stream of gene

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Step 1 Denaturing
60 seconds _at_ 94C
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Step 2 Annealing
60 seconds _at_ 60C Forward and Reverse Primers
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Taq Polymerase Binds
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Step 3 Extension
2 minute at 72C DNTPs
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PCR Flash Animation
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(No Transcript)
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The exponential amplification of the gene in PCR.
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Amplification
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Cycle 3

• Obtain two copies of target DNA

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Cycle 30

• 1,073,741,764target copies

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Is there a gene copied during PCR and is it the
right size ?
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Applications of PCR

• quick, reliable method for detecting all manner
  of mutations associated with genetic disease -
  from insertions, to deletions, to point
  mutations.
• Duchene muscular dystrophy
• Detect unwanted Genetic material
• Bacterial or viral infection
• HIV infection
• Amplify degraded DNA samples
• Egyptian mummy
• Termite in amber

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PCR animation
PCR animation
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On-Line Sources

• Access Excellence. Polymerase Chain Reaction
  (PCR) 1992.19 Oct 2004. lthttp//www.accessexcelle
  nce.org/RC/VL/GG/polymerase.htmlgt.
• Access Excellence. Polymerase Chain Reaction -
  From Simple Ideas 1999.19 Oct 2004.
  lthttp//www.accessexcellence.org/RC/AB/BC/PCR_From
  _Simple_Ideas.htmlgt.
• Access Excellence. Polymerase Chain Reaction -
  Xeroxing DNA 1992.19 Oct 2004.
  lthttp//www.accessexcellence.org/RC/AB/IE/PCR_Xero
  xing_DNA.htmlgt.
• Cold Spring Harbor Laboratory.1994 Dolan DNA
  Learning Center. 18 Oct 2004.lthttp//www.dnalc.org
  /resources/BiologyAnimationLibrary.htmgt.
• Montpetit, Diane. Electron microscopy 17Nov
  2003. Agriculture and Agri-Food Canada. 19 Oct
  2004. lthttp//www.agr.gc.ca/disc_e.phtmlgt.
• Nobelprize.org. The Nobel Prize in Chemistry
  1993. 1 Nov2000. lthttp//nobelprize.org/chemistry/
  laureates/1993/index.htmlgt.
• Nobelprize.org. The PCR Method - a DNA Copying
  Machine. 19June2004. lthttp//nobelprize.org/chemis
  try/educational/pcr/index.htmlgt.
• Vierstraete, Andy. Principle of the PCR
  08-11-1999. University of Ghent.19 Oct 2004
  lthttp//allserv.rug.ac.be/7Eavierstr/principles/p
  cr.htmlgt.
• Ward, Laura. Food for thought 2004 LABORATORY
  NEWS magazine. 19 Oct 2004lthttp//www.labnews.co.u
  k/features_5.htmgt.

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Book Sources

• Micklos, David, Greg Freyer and David Crotty. DNA
  Science a First Course. New YorkCold Spring
  Habor Laboratory Press, 2003.