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Molecular cloning

Cloning: Selection

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Learn how antibiotic resistance can be used to select for bacteria that have taken up plasmid DNA, and about the key stages in the plasmid selection process. Practise interpreting results from sample agar plates and cloning experiments.

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Learning outcomes

  • Predict the cells you would expect to find growing in a recovery culture when transformation is complete
  • Understand how antibiotic resistance can be used to select for bacteria containing plasmids
  • Interpret the results of cloning experiments including the use of controls
  • Transform competent cells with recombinant plasmid
  • Select for cells that have taken up plasmid
  • Confirm that the cloning process was successful
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Skills

Designing and Planning Experiments

  • Why the selectable marker in the plasmid determines the choice of antibiotic used in the agar 
  • Why an L-spreader is the most appropriate instrument to spread culture on agar 
  • How use of a positive control can validate the transformation procedure 
  • How use of a negative control can validate the selection procedure

Interpreting and Analysing Data

  • How to use the results of sample and control plates to identify sources of error in the procedure

Concepts, Principles and Theories

  • How bacterial transformation can produce bacteria with/without the plasmid and with/without the desired insert
  • What is happening on a molecular level when transformed cultures are incubated on agar containing antibiotic

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Details

Degrees

Biomedical science
Biological science
Pharmacological science
Forensic science
Biochemistry

Modules

Molecular biology & genetics

Topic

Molecular cloning

Purchase option

Bioscience LabSim Library

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