Skills in Molecular Bioscience

Learning Outcomes

Students will learn about current research techniques that are used widely in research laboratories in order to gain insight into gene expression and protein interactions. Each work package will require students to engage with a key technique through lectures and workshops, and then produce a piece of assessed work which demonstrates that they can design experiments or analyse data. Work packages will cover the following topics: 1) Gene cloning – how do I clone gene X or Y from cDNA? We will consider the source of appropriate starting material (e.g. tissue/cell type), primer design, vector choice, and how to undertake classic genetic engineering. Activities: Design PCR primers to amplify the ORF of a gene of interest, and determine how to clone this into vectors containing epitope tags or fluorescent proteins. 2) Gene manipulation (qPCR and siRNA). Discussion of the background and applications of these techniques. How to design a knockdown specific to your gene of interest? How to measure RNA levels following knockdown? Activities: design of siRNA targets, and then a vector to escape any phenotype associated with siRNA-mediated knockdown. You will also design qPCR primers to analyse levels of expression of your gene of interest (or the human homolog). You will analyse qPCR data to determine primer amplification efficiency, and relative expression levels of the gene. This data has to be presented appropriately, with statistical tests applied. 3) Protein domains – what are the functions of my protein? Activities: Design an experiment to mutate a domain/site of interest in the plasmid constructed in work package 1. 4) Gene manipulation (CRISPR). Discussion of background and applications. Activities: Give oral presentation on your protein of interest discussing domains and sites of interest and how you would plan an experiment to edit in amino acid mutations, or to add an epitope tag to your protein. Submit documentation on how you would: design guides for insertion into a Cas9 plasmid; design the template for homology directed repair; design the experiment for genotyping the knock-in.

Each work package will begin with a lecture on a molecular biology technique, and this will be followed up with a workshop where students will use electronic resources to design experiments.