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Molecular Cloning

Blue-White Screening Theory

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Learn the molecular basis of blue-white screening, including the role of the lac operon, beta-galactosidase and alpha-complementation. Explore how the success of ligation and transformation steps affects the growth and appearance of bacteria on the plate and practise identifying colonies containing recombinant bacteria.

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Learning Outcomes

  • Explore the function and molecular basis of blue-white screening
  • Determine the role of each reagent
  • Explain key results in terms of transformation success
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Skills

Obtaining and Recording Data

  • Why the agar must be supplemented with the appropriate antibiotic, IPTG and X-gal

Interpreting and Analysing Data

  • How to identify colonies that have taken up the plasmid with the desired insert

Concepts Principles and Theories

  • What is the purpose of blue-white screening
  • What components make up the lac operon in E. coli
  • Why IPTG is seen as an activator of the lac operon
  • Why bacteria that have not taken up the plasmid die before forming colonies
  • Why bacteria that have taken up the plasmid with no insert will form blue colonies
  • Why bacteria that have taken up the plasmid with insert will form white colonies

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Student, Natural Sciences
University of Exeter

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Details

Degrees

Biochemistry
Biomedical Science
Biological Science
Pharmacological Science
Forensic Science

Modules

Molecular Biology & Genetics

Topic

Molecular Cloning

Purchase option

Bioscience LabSim Library

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