The University of Southampton
Courses

BIOL6077 Skills in Molecular Bioscience

Module Overview

Aims and Objectives

Module Aims

Aims and Learning Outcomes ________________________________________ 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 interaction studies. Critically evaluate experimental techniques used in the wider molecular bioscience field.

Learning Outcomes

Learning Outcomes

Having successfully completed this module you will be able to:

  • 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 TOPO versus classic genetic engineering. Activities: make virtual plasmids for E. coli expression/purification of a gene of interest, and also create virtual vectors for mammalian expression of your protein of interest fused to epitope tags or fluorescent proteins. 2) qPCR – How to measure RNA levels, include some discussion of RNA sample preparation Activities: design of experiment for determining expression levels genes of interest, including the use of appropriate controls. Analysis of provided data using both the delta Ct method and standard curve-based. This has to be presented appropriately, with statistical tests applied. 3) Protein assays – antibody design, protein interaction studies (incl proteomics) Activities: Analyse proteomic output (using InTact from EMBL/EBI) and make interaction map (STRING) and analyse pathways using DAVID. 4) Gene manipulation (siRNA). Discussion of the background and applications of the technique. Activities: design targets, and then a vector to escape any phenotype associated with siRNA-mediated knockdown (Quikchange method). You will also design qPCR primers to analyse knockdown of expression. 5) 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.

Syllabus

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.

Learning and Teaching

TypeHours
Independent Study60
Teaching15
Total study time75

Assessment

Summative

MethodPercentage contribution
Coursework 20%
Coursework 20%
Coursework 20%
Coursework 20%
Practical exercise 20%
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