This module aims to provide the student with: An understanding of and competency in basic polymerase chain reaction (PCR), recombinant DNA technology, A knowledge of the basic concepts of mammalian cell culture; An understanding of the principles of protein isolation and purification, and the necessary skills to isolate and characterise proteins; immunoassays and their role in protein analysis An understanding of protein diseases and of the roles played by proteins in medicine; Theoretical aspects of this module are complemented by laboratory experiments delivered in the Laboratory Skills module running concurrently.
DNA Cloning
DNA isolation and analysis; restriction and ligation of DNA into a plasmid vector, transformation of recombinant plasmid into prokaryotic cell; basic concepts of polymerase chain reaction
Basic Mammalian Cell Culture
Introduction to cell culture; primary cells and cell lines; Adherent and Suspension cells; subculturing of mammalian cells; safety
Proteins in Medicine and Disease:
The role played by proteins in medicine. Therapeutic proteins; Types of protein - related diseases, e.g. Prion proteins; cystic fibrosis; Methods of prevention and protection.
Protein Assays:
Types of protein assays; Immunoassays; Sensitivity; Choosing an assay; Miniaturisation
Protein Purification:
Physical, chemical and functional features which may be exploited in purification (e.g. post translational modification); Sample preparation;Extraction from cells; Solubilisation of membrane proteins; Immuno-precipitation; Protein concentration - Salting out; TCA precipitation; Ultrafiltration;
Electrophoresis:
Quantitation; Characterisation (Size / Subunits / Sequence); Factors affecting migration; Continuous and Discontinuous buffer systems; IEF and SDS electrophoresis; MW Determinations; Western Blotting.
Chromatography:
Theory, principles and applications approaches to protein purification – Size-exclusion / Ion-exchange / gel filtration / Affinity / HPLC chromatography
| Module Content & Assessment | |
|---|---|
| Assessment Breakdown | % |
| Formal Examination | 70 |
| Other Assessment(s) | 30 |