Polymerase Chain Reaction

1. Polymerase Chain Reaction

2. PCR... Whaaaat? Founded by Kary Mullis in 1984 He wished to bracket the desired section of DNA with primers and copy the desired fragment with DNA polymerase. Essentially “Genetic Photocopying”

3. Besides DNA, there are 3 main ingredients:: The “Deets” Ta Primers Taq polymerase --> High Temperatures  Free nucleotides • Cycle 1 • A temperature "yoyo" • Taq polymerase --> 3' - 5'

4. The "Deets" (cont'd) Cycle 2  - repeat cycle 1 :) - 4 copies of target DNA After 32 cycles of PCR there will be: 4 294 967 296 copies of target DNA  Cycle 3  - exactly like cycles 1 and 2 - 8 copies of target DNA Note** only 2 of these copies are of the target fragment on its own.   

5. What the Cycles Look Like

6. Works Cited “Polymerase Chain Reaction.” Contexo.info. Web Page. 19 Nov 2010. • http://www.contexo.info/DNA_Basics/polymerase_chain_reaction.htm. • “PCR.” Enigma Diagnostics. 2009. Web Page. 28 Nov 2010. • http://www.enigmadiagnostics.com/template2.php?page=technology.php&m=2. • “The Polymerase Chain Reaction.” Sumanasinc.com. Web Page. 19 Nov 2010. • http://www.sumanasinc.com/webcontent/animations/content/pcr.html. • “Polymerase Chain Reaction.” Genetics Wiki. 30 Sept 2009. Web Page. 19 Nov 2010. • http://wiki.cstl.semo.edu/agathman/PCR.ashx • “Scientist Develops Salmonella Test That Makes Food Safer, Reduces Recalls.” Meat • and Poultry Online. 24 Nov 2010. Web Page. 25 Nov 2010. • http://www.meatandpoultryonline.com/article.mvc/Scientist-Develops- Salmonella-Test-0001?atc~c=771+s=773+r=001+l=a&VNETCOOKIE=NO. • “Using molecular marker technology in studies on plant genetic diversity: DNA-based • technologies; PCR based technologies: PCR basics. Cornell University. 2003. • Web Page. 28 Nov 2010. • www.bioversityinternational.org/.../MolMarkers%20Vol1%20III%20PCR%20basics.pdf. • “What is a Thermal Cycler.” Thermal Cycler. Web Page. 28 Nov 2010. • http://thermalcycler.net/.

7. 3 Major Steps Each Cycle of Polymerase Chain Reaction can be split into 3 basic steps:          Step 1: Denaturation

8. Step 2: Annealing

9. Step 3: Extension

10. Further Explanation http://www.sumanasinc.com/webcontent/animations/content/pcr.html

11. Heavy Machinery Thermal Cycler is used in the lab to make large numbers of DNA fragments

12. Look Familiar Much? DNA replication and PCR are very similar....they both: need Template stands Needs polymerase enzyme Both use primers

13. Applying PCR to the Real World Human Genome Project DNA Fingerprinting / Forensics Mutation Screening Genealogical Testing / Genetic Disorders Cloning Detection of viruses (namely HIV and AIDS) ... Just to name a few

14. PCR in the NEWS! • Scientist at University of Missouri using PCR to detect salmonella in eggs and poultry

15. THE END And now for your listening pleasure… http://www.cnpg.com/video/flatfiles/539/