Upload
reynard-lawrence
View
214
Download
0
Embed Size (px)
Citation preview
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”
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'
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 DNANote** 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
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/.
3 Major Steps
Each Cycle of Polymerase Chain Reaction can be split into 3 basic steps:
Step 1: Denaturation
Look Familiar Much?
• DNA replication and PCR are very similar....they both:
• need Template stands• Needs polymerase enzyme• Both use primers
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
PCR in the NEWS!
• Scientist at University of Missouri using PCR to detect salmonella in
eggs and poultry