Comprehensive Notes for BCMB20005 Exam (H1)

Description

Includes all topics covered in lectures: - Pipetting technique & spectrophotometry - UV analysis of DNA & proteins - Basic gel electrophoresis technique, as well as native gel electrophoresis, SDS-PAGE and vertical acrylamide gel electrophoresis - Function and application of molecular tools including restriction enzymes and DNA ligase - Southern and Western Blot techniques - Basic PCR technique, as well as rtPCR and qPCR (Sybr and Taqman assays) - Process of bacterial transformation with a gene of interest via both heat shock and electroporation - Process of bacterial selection via antibiotic resistance and blue/white selection - Features of cloning and expression plasmids - Construction and purpose of both genomic and cDNA libraries - Cell biology and application of both adherent and suspension culture - Basic Michaelis Menten enzyme kinetics and association with GLUT transporters - Acid-base chemistry and application to proteins via fractional charge calculation - Protein assays - Biuret, Bradford, Lowry - and their parameters and applications - Bacterial expression systems (pET & pGEX) and expression via the lac Operon - Purification of proteins via dialysis or ultrafiltration - Purification of proteins via chromatography - ion exchange chromatography, size exclusion chromatography and affinity tag chromatography - Precipitation techniques including pH, salt and immunoprecipitation - Protein purification tables - Other eukaryotic expression systems including baculovirus and yeast plasmids (YEp & YIp)