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Polymerase Chain Reaction
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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”
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The “Deets” Ta Primers Taq polymerase --> High Temperatures Free nucleotides Besides DNA, there are 3 main ingredients :: Cycle 1 A temperature "yoyo" Taq polymerase --> 3' - 5'
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The "Deets" (cont'd) Cycle 2 - repeat cycle 1 :) - 4 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. After 32 cycles of PCR there will be: 4 294 967 296 copies of target DNA
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What the Cycles Look Like
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Works Cited “Polymerase Chain Reaction.” Contexo.info. Web Page. 19 Nov 2010. http://www.contexo.info/DNA_Basics/polymerase_chain_reaction.htm.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.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.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.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.www.bioversityinternational.org/.../MolMarkers%20Vol1%20III%20PCR%20basics.pdf “What is a Thermal Cycler.” Thermal Cycler. Web Page. 28 Nov 2010. http://thermalcycler.net/.http://thermalcycler.net/
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3 Major Steps Each Cycle of Polymerase Chain Reaction can be split into 3 basic steps: Step 1: Denaturation
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Step 2: Annealing
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Step 3: Extension
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Further Explanation http://www.sumanasinc.com/webcontent/animations/content/pcr.html
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Heavy Machinery Thermal Cycler is used in the lab to make large numbers of DNA fragments
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Look Familiar Much? DNA replication and PCR are very similar....they both: need Template stands Needs polymerase enzyme Both use primers
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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
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PCR in the NEWS! Scientist at University of Missouri using PCR to detect salmonella in eggs and poultry
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THE END And now for your listening pleasure… http://www.cnpg.com/video/flatfiles/539/
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