Aims & Learning Objectives:
Aims: To provide practical experience of some commonly used molecular biology techniques and to evaluate these in comparison with other available methods so that the students are aware of a range of techniques used for the characterisation of gene structure and gene expression.
After taking this course the student should be able to:
* understand how and why these techniques have been applied when they encounter them in journal articles
* consider these methods when they design their own experiments
* develop planning and organisational skills in carrying out a scientific project
* develop data handling and interpretation skills
Content:
The course involves a series of linked experiments that form a mini-project. This begins with the preparation of cellular RNA, its conversion to cDNA and the amplification of gene fragments using the polymerase chain reaction (PCR). Specific fragments are cloned in a bacterial plasmid, these screened by restriction enzyme digestion and one or two are subject to DNA sequencing. Data interpretation involves DNA sequence analysis and the interrogation of remote nucleic acid databases, chromosome physical maps and mutational databases. A demonstration of various methods for analysing gene expression is included.
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