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Polymerase Chain Reaction (PCR)

Understanding qPCR

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Learn about incorporating quantification in PCR. Explore the principles of fluorescence and two different fluorescence detection methods used in qPCR. Understand the progression of a standard amplification plot and review a set of qPCR results.

Learning Outcomes

  • Review the fundamentals of PCR and its quantification
  • Identify the principles and differences of two fluorescence methods
  • Recognise the progression of a standard amplification plot
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Designing and Planning Experiments

  • How SYBR green and TaqMan probes differ in their specificity for DNA, interaction with DNA and cost
  • What are some of the key differences between endpoint PCR and qPCR

Interpreting and Analysing Data

  • What axes are used for an amplification plot 
  • How does the curve on an amplification plot reflect what is happening on a molecular level in the sample
  • How to use an amplification plot to compare the starting quantities of target DNA

Concepts Principles and Theories

  • What are the 3 main stages of a PCR cycle and the temperatures at which they occur
  • What is meant by the term fluorophore
  • Why an increase in target DNA proportionally increases the fluorescence by SYBR green
  • Why an increase in target DNA proportionally increases the fluorescence by TaqMan probes 
  • Why the process is called qPCR or RT-PCR

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Polymerase Chain Reaction (PCR)

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