Skills in Molecular Bioscience

Learning Outcomes

Learning Outcomes

Having successfully completed this module you will be able to:

• Having successfully completed this module, you will have developed skills in different areas of molecular bioscience: 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) Protein assays – what are the functions of my protein? Activities: Give an oral presentation on your protein of interest, discussing domains and sites of interest. Design an experiment to mutate a domain/site of interest and hypothesise likely experimental outcomes. 3) Gene manipulation (siRNA and qPCR). 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 knockdown of expression. Analysis of provided data using the delta Ct method. This has to be presented appropriately, with statistical tests applied 4) Gene manipulation (CRISPR). Discussion of background and applications. Activities: Design guides for WT and nickase plasmids, design gBLOCK for HDR (including checking restriction sites/cloning into HDR template plasmid), design experiment for genotyping.

Knowledge and Understanding

Having successfully completed this module, you will be able to demonstrate knowledge and understanding of:

• By the end of this module students will be able to: Understand and demonstrate the planning of several different experimental techniques in molecular biology. Analyse and present data from gene expression and protein characterisation studies. Critically evaluate experimental techniques used in the wider molecular bioscience field.

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.

Summative

This is how we’ll formally assess what you have learned in this module.