Lecture 5 - BIO 311 Polymerase Chain Reaction Fall 2010...

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BIO 311 Polymerase Chain Reaction Fall 2010
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Amplifying DNA in Vitro : The Polymerase Chain Reaction (PCR) The polymerase chain reaction, PCR , can produce many copies of a specific target segment of DNA A three-step cycle—heating, cooling, and replication—brings about a chain reaction that produces an exponentially growing population of identical DNA molecules
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Fig. 20-8 5 Genomic DNA TECHNIQUE Cycle 1 yields 2 molecules Denaturation Annealing Extension Cycle 2 yields 4 molecules Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence Target sequence Primers New nucleo- tides 3 3 3 3 5 5 5 1 2 3
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Fig. 20-8a 5 Genomic DNA TECHNIQUE Target sequence 3 3 5
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Fig. 20-8b Cycle 1 yields 2 molecules Denaturation Annealing Extension Primers New nucleo- tides 3 5 3 2 5 3 1
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Fig. 20-8c Cycle 2 yields 4 molecules
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Fig. 20-8d Cycle 3 yields 8 molecules; 2 molecules (in white boxes) match target sequence
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PCR’s History The Discovery of: The Polymerase Chain Reaction (PCR) It’s the story of an “Unconventional” Scientist Dr. Kary B. Mullis Described as: “An “intellectual maverick”” “One of the most unusual scientists of our times”
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Where and When? May 1983 - Mullis invented the Polymerase Chain Reaction (PCR) He was driving to Anderson Valley on California’s Highway 128 with Jennifer – his girlfriend and co-worker He was thinking about…
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A Chain Reaction “A series of events so related to each other that each one initiates the next” “ A number of events triggered by the same initial event”
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Making PCR Work Mullis took his concept to his associates at Cetus Together they took the idea and made it work in an experimental system Mullis received a $10,000 bonus for his idea of PCR 1992- Cetus sold the patent to Hoffman-LaRoche for $300 million dollars!
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A Nobel Laureate Mullis was awarded both the Japan Prize and the Nobel Prize in Chemistry in 1993 Mullis lectures around the world about biotechnology or the development of the scientific method
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Polymerase Chain Reaction A common misconception : This is simply NOT the case… PCR = (For DNA)
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So…What exactly is PCR? Polymerase Chain Reaction (PCR) : “A procedure that enzymatically amplifies a target DNA sequence of up to several thousand base pairs through repeated replication by DNA polymerase” - DNA Science (Micklos & Freyer) To understand how we direct it to amplify a given site (and not some other site) we must first learn the mechanism! Though not a Textbook definition, PCR Can be thought of as a tool that allows for the Site Directed Amplification of Template DNA (up to around 10 KB in size)
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Copy Machine vs. PCR Standard Copy Machines copy all the data put into it (i.e. the entire page) PCR “copies” only what you design it to
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Copy Machine vs. PCR
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Lecture 5 - BIO 311 Polymerase Chain Reaction Fall 2010...

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