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Polymerase Chain Reaction (PCR)

Running a Polymerase Chain Reaction (PCR)

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Learn how to set up a PCR reaction by preparing four tubes for an experiment, including a positive and negative control. Explore the steps of adding dNTPs, primers, samples and enzymes to each tube, and set the thermocycler to the correct settings. Analyse the results by running samples on an agarose gel and interpreting the outcome.

Learning Outcomes

  • Add key reagents of a PCR experiment to four PCR tubes and consider their function
  • Set up negative and positive controls
  • Analyse the results of the PCR by studying an agarose gel of the amplified DNA
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Designing and Planning Experiments

  • Why dNTPs, DNA, primers and polymerase enzyme are required in each sample tube 
  • Where primers should be targeted in relation to the region of interest
  • What makes an appropriate positive and negative control in a PCR experiment

Using Instruments and Equipment

  • How to set the desired cycle settings on the thermocycler

Obtaining and Recording Data

  • Why it is important to prepare PCR samples on ice 
  • Why it is best to add the DNA polymerase to the reaction mix last

Interpreting and Analysing Data

  • How to use an agarose gel to identify whether samples and controls contain the region of interest

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Polymerase Chain Reaction (PCR)

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