Project Title: The Effects of Using Lysate from Acid-Stressed E. coli on the Yield and pH homeostasis of Cell-Free Protein SynthesisSupervisor: Dr Antonios BougasOverview: Producing call lysates from acid-stressed and non-stressed E. coli to be used in comparative cell-free protein synthesis (CFPS) reactions. Lysate was produced from E. coli cultured using a self-developed protocol. This was used to investigate the effect of acid-stressed lysate on the protein yield and pH homeostasis of CFPS reaction in the presence and absence of conventional pH buffers with the hypothesis that acid-stressed lysates may be able to self regulate CFPS reaction pH in the absence of conventional pH buffers. This was a self arranged project that built on the experience and results produced during my iCM internship at the Regan lab. Technical Skills: bacterial cell culture, CFPS reaction assembly, plasmid purification, experiment design, data analysis, oral presentation

Overview: During this internship I investigated different methods of increasing the yield of E. coli based cell-free protein synthesis reactions. This included the addition and removal of reaction additives as well as by altering the proportions of key reaction components (DNA template, energy solution, bacterial lysate). The results and experience from this internship formed the basis of my self-organised honours research project-Supervisor:Dr Ella Thornton, The Regan Lab, The University of Edinburgh. Technical Skills: CFPS reaction assembly, PCR reaction assembly, gel electrophoresis, plasmid DNA purification, independent research, data analysis and presentation