1. PCR Polymerase Chain Reaction Revised 10-31-2006

2. 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”

3. Taq polymerase

4. Thermus aquaticus

5. Thermal Cycler

6. Materials of PCR Materials of PCR target DNA target DNA Taq DNA polymerase Taq DNA polymerase 2 Primers 2 Primers ~20 nucleotides in length ~20 nucleotides in length Forward and reverse Forward and reverse the four DNTP’S the four DNTP’S Adenine Adenine Thymine Thymine Cytosine Cytosine Guanine Guanine cofactor MgCl 2. cofactor MgCl 2.

7. Locate the Target Sequence Scientists determine which GENE they are interested in studying Scientists determine which GENE they are interested in studying Locate Primers Upstream and Down stream of gene Locate Primers Upstream and Down stream of gene

8. Step 1 Denaturing 60 seconds @ 94°C

9. Step 2 Annealing 60 seconds @ 60°C Forward and Reverse Primers

10. Taq Polymerase Binds

11. Step 3 Extension 2 minute at 72°C DNTP’s

12. PCR Flash Animation

14. The exponential amplification of the gene in PCR.

15. Amplification

16. Cycle 3 Obtain two copies of target DNA Obtain two copies of target DNA

17. Cycle 30 1,073,741,764 target copies 1,073,741,764 target copies

18. Is there a gene copied during PCR and is it the right size ?

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

20. PCR animation

21. On-Line Sources Access Excellence. “Polymerase Chain Reaction (PCR)” 1992.19 Oct 2004.. http://www.accessexcellence.org/RC/AB/BC/PCR_From_Simple_Ideas.html Access Excellence. “Polymerase Chain Reaction - From Simple Ideas” 1999.19 Oct 2004.. Access Excellence. “Polymerase Chain Reaction - Xeroxing DNA ”1992.19 Oct 2004..http://www.accessexcellence.org/RC/AB/IE/PCR_Xeroxing_DNA.html http://www.dnalc.org/resources/BiologyAnimationLibrary.htm Cold Spring Harbor Laboratory.1994 Dolan DNA Learning Center. 18 Oct 2004... 19 Oct 2004.. Montpetit, Diane. “Electron microscopy” 17Nov 2003. Agriculture and Agri-Food Canada. 19 Oct 2004.. Nobelprize.org. The Nobel Prize in Chemistry 1993. 1 Nov2000..http://nobelprize.org/chemistry/laureates/1993/index.html Nobelprize.org. The PCR Method - a DNA Copying Machine. 19June2004.. Vierstraete, Andy. “Principle of the PCR” 08-11-1999. University of Ghent.19 Oct 2004.http://allserv.rug.ac.be/%7Eavierstr/principles/pcr.html Ward, Laura. “Food for thought” 2004 LABORATORY NEWS magazine. 19 Oct 2004. http://www.labnews.co.uk/features_5.htm

22. Book Sources Micklos, David, Greg Freyer and David Crotty. DNA Science a First Course Micklos, David, Greg Freyer and David Crotty. DNA Science a First Course. New York:Cold Spring Habor Laboratory Press, 2003.