Molecular Biology
Overview
- Credit value: 15 credits at Level 5
- Convenor: Dr Tracey Barrett
- Assessment: lab reports, blog compilation, short-answer final test (100%); 80%Â attendance at lab-based sessions (0%)
Module description
In this module you will be provided with a robust 'toolkit' of laboratory techniques and approaches within the realm of recombinant DNA technology, including PCR, DNA assembly/cloning and DNA sequencing. All of this will build on the relevant knowledge and practical skills acquired at Level 4 and in the laboratory sequence within the Level 5 module, Evolution and Genetics.
The knowledge and experience you will gain in this module will serve as essential preparation work at Level 6, especially in the final-year laboratory research projects.
Indicative syllabus
- PCR: principles; key applications; primer design
- DNA polymerases: choices for PCR and other applications
- DNA restriction: applications in mapping and cloning
- Plasmids and other cloning vectors
- DNA intermolecular ligation and transformation into competent cells
- Sanger fluorescent DNA sequencing analysis
Learning objectives
By the end of this module, you will be able to:
- explain the general principles of gene cloning and identify key considerations in regard of choice of cloning vector
- explain in general how the polymerase chain reaction (PCR) works and give examples of how PCR may be applied in DNA cloning and sequencing workflows
- design appropriate primers and explain how you would know if the PCR had been successful when given a target DNA sequence for amplification by the polymerase chain reaction
- give details of the Gibson assembly process for DNA fragment assembly, describe how the fragments obtained may be introduced into a plasmid and explain how you would determine if the method had been successful
- describe techniques for the introduction of recombinant DNA into cells, and explain methods for the downstream isolation of this DNA and for determination of the quality of the product
- outline the workflow leading to Sanger sequencing, explain how the technique works and interpret the readout generated by this process.