PCR and Restriction Cloning

This lecture covers the genetic engineering and characterization of a FRET-sensor of enzymatic activity, focusing on topics such as PCR cloning, primer design, restriction & ligation, good FRET pairs, electrophoresis, and the design of DNA constructs. The instructor explains the protease cleavage site, the FRET pair, and the conditions for a good FRET sensor. Additionally, the lecture delves into plasmids, restriction endonucleases, DNA ligases, and the polymerase chain reaction process. Practical aspects, such as electrophoresis and DNA sequence analysis, are also discussed, along with the necessary preparations for the laboratory sessions.