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Mastering PCR: Principles, Techniques & Troubleshooting Guidelines

Learn the fundamentals and troubleshooting tips for Polymerase Chain Reaction (PCR) in this comprehensive workshop. Understand the basics of PCR, exponential amplification rates, and how to properly quantify products. Troubleshoot common PCR issues effectively.

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Mastering PCR: Principles, Techniques & Troubleshooting Guidelines

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  1. Principles of PCR MUPGRET Workshop June 18, 2004

  2. PCR • Polymerase Chain Reaction • http://www.dnai.org/b/index.html • http://palou.uib.es/roolpi/PCR/

  3. Basic PCR • Initially reagents in excess, template and product don’t renature and outcompete primer binding, so amplification occurs at an exponential rate. • Timepoint when exponential turns to linear rate is different depending on the initial concentration of the target molecule.

  4. Basic PCR • To properly quantify one product relative to another you need to be in the exponential phase of amplification for both and know when the end point occurs.

  5. Troubleshooting PCR • http://www.promega.com/guides/pcr_guide/070_22/promega.html • http://info.med.yale.edu/genetics/ward/tavi/Trblesht.html • http://palou.uib.es/roolpi/PCR/

  6. Electrophoresis • A way to separate DNA molecules. • http://gslc.genetics.utah.edu/units/biotech/gel/

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