1. Real Time PCR as a Diagnostic Tool

2. 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=sGP 56W0CSV8

3. 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

4. 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.

5. What’s a Single Nucleotide Polymorphism (SNP)? Do the tutorial at http://learn.genetics.utah.edu/content/health/pharma/snips/

6. 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.

7. Lightcycler data