Polymerase Chain Reaction

1
Polymerase Chain Reaction

• Core 218 Biotechnology and Society
• Lecture 15
• Spring 2005

2
The Polymerase Chain Reaction

• PCR Polymerase Chain Reaction, a method to
  amplify fragments of DNA in vitro.
• DNA Fingerprinting
• Diagnosis of inherited disorders
• Developing cures for inherited disorders
• Biological evidence
• Personal identification

3
PCR Conception

• Kary B. Mullis
• Education
• B.S. Chemistry from Georgia Tech
• Ph.D. in Biochemistry from U.C. Berkeley.
• His Ph.D. thesis was entitled, "Schizokinen
  Structure and Synthetic Work describing a
  molecule involved in iron transport.
• The Cosmological Significance of Time Reversal.
  Nature 218663 ( 1968).

4
Kary Mullis

• Cetus Corporation, Emeryville in the 1970s.
• Honda Civic on Highway 128 from San Francisco to
  Mendocino
• Nobel Prize in Chemistry in 1993 for developing
  PCR
• 10,000 from Cetus
• Currently, he consults and lectures around the
  world about cosmology, mysticism, mathematics,
  virology, and artificial intelligence

5
The PCR Reaction

• Template DNA
• Primer
• Polymerase from Thermus aquaticus
• Innis, M.A., K.B. Myambo, D.H. Gelfand, and M.A.
  Brow. (1988) DNA sequencing with Thermus
  aquaticus DNA polymerase and direct sequencing of
  polymerase chain reaction-amplified DNA. Proc.
  Natl. Acad. Sci. USA85 9436
• Nucleosides (A, T, G, C)
• Hi and low temperature reactions
• 90-95 C, strand separation
• 55 C, primer reanneals
• 75 C, Taq polymerase synthesizes new strand
• Repeat temperature cycle at least 30 times

6
The PCR Reaction
http//allserv.rug.ac.be/avierstr/principles/pcra
ni.html