real time pcr as a diagnostic tool. pcr = polymerase chain reaction a way to make lots of copies of...
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![Page 1: Real Time PCR as a Diagnostic Tool. PCR = Polymerase Chain Reaction A way to make lots of copies of DNA What Happens in the Machine? Do the PCR virtual](https://reader035.vdocument.in/reader035/viewer/2022062715/56649d965503460f94a7e8ff/html5/thumbnails/1.jpg)
Real Time PCRReal Time PCRas a Diagnostic as a Diagnostic
ToolTool
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PCR = Polymerase Chain Reaction• A way to make lots of copies of DNA
What Happens in the Machine?Do the PCR virtual lab at
http://learn.genetics.utah.edu/content/labs/pcr/
Setting Up!See real people doing PCR at
http://www.youtube.com/watch?v=sGP56W0CSV8
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Real Time PCR
• Amplifies AND quantifies a targeted segment of DNA– Regular PCR: DNA detected at end– Real time PCR: DNA detected as the reaction
progresses• Fluorescent dyes or probes are added and
fluorescence is measured during the reaction• Can be used for both diagnostic & basic
research
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The Lightcycler (Real-time PCR)
• Used to detect SNPs- single nucleotide polymorphisms- based on melting curve analysis
• Quick, inexpensive way to investigate genetic differences; works as a diagnostic tool.
The Lightcycler consists of a Computer attached to a PCR machine with a florescence detector.
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What’s a Single Nucleotide Polymorphism (SNP)?
Do the tutorial at http://learn.genetics.utah.edu/content/health/pharma/snips/
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Melting Curve Analysis
• The temperature at which 50% of the DNA is denatured is called the melting point.– G-C pairs have 3 hydrogen
bonds.– A-T pairs have 2 hydrogen
bonds.– G-C bonds will “melt” at a
higher temperature.
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Lightcycler data