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Molecular Cloning

Plasmid Ligation

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Learn about the catalysis of a ligation reaction by DNA ligase, and explore the benefits of using phosphatase treatment and double digests to prevent the religation of plasmids.

Learning Outcomes

  • See how DNA ligase catalyses a ligation reaction
  • Consider the benefits of using phosphatase treatment before ligation
  • Look at the use of double digests to prevent plasmid religation
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Designing and Planning Experiments

  • What reagents and cofactors are required for the ligation reaction 
  • How to select the most suitable restriction enzymes for a double digest

Concepts Principles and Theories

  • What is happening on a molecular level during the digestion and ligation reaction 
  • How the use of phosphatase or multiple restriction enzymes can prevent religation of the plasmid DNA

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Molecular Cloning

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