BIOU7TM: Molecular Techniques

CO-ORDINATOR: Dr Mike Wyman
CONTRIBUTORS: Dr Tim Whalley

Aims and Objectives

The main aims of the Molecular Techniques half module are to provide practical experience in:

  • cloning techniques
  • the over-expression of a cloned gene
  • the purification of an over-expressed protein
  • localisation of protein in vivo

This half module provides opportunities to develop your knowledge and understanding of modern molecular techniques.

Acquired Skills

  • Knowledge: developing background knowledge in molecular techniques
  • Written communication: Report writing and note taking
  • IT: Data analysis and presentation
  • Information retrieval: use of library and internet
  • Time management: covering course material, practical work and report preparation

Assessment

Lab report: 3,500 words (100%)

Lab Classes

Lab classes will be in 2U2

Practical Programme

  • Induction of GFP overexpression in E. colI
  • GFP purification and spectroscopic analysis
  • Transformation of  S.  pombe with GFP labelled calnexin construct
  • Localisation of calnexin- GFP using microscopy

Recommended Reading

  • Exploring Proteins (2009) Price and Nairn
  • Molecular Cell Biology 6th Edition (2007) (Chapter 5; Molecular Genetic Techniques) Lodish et al
  • Biology 8th Edition (2007) (Chapter 18; Gene Expression) Campbell & Reece

Report Guidelines

Report should be prepared with the following headings:

Abstract, Introduction, Methods and Materials (simply refer to protocol i.e. no need to rewrite protocol), Results, Discussion, Conclusion and References

The introduction should provide some background on the applications of protein overexpression and GFP as a tool probe cell structure/function.

Your discussion should be clear, concise and relate your results to appropriate published work. References should be from peer reviewed journals i.e. do not produce a list of websites

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