BCMB20005 Lecture Notes - Lecture 2: Gc-Content, Antimicrobial Resistance, Filter Paper

• School: University of Melbourne
• Department: Biochemistry and Molecular Biology
• Course: BCMB20005
• Professor: Amber Willems

Document Summary

Isolation and analysis of plasmid dna steps involved in the isolation. Restriction enzymes and the manipulation of dna mode of action and what the mechanism behind the hydrolysis of phosphodiester bond. Southern blot analysis steps involved and the dna hybridization properties such as temperature, including tm. Step 1: isolate dna from either plants or animals. Plasmids can replicate independently, hence very useful in cloning vector. Useful in transferring, manipulating and replicating dna sequences, as desired genes can be inserted into the circular plasmid, and manipulated to create more copies. Step 2: obtain dna fragment of interest: use restriction enzymes (restriction endonuclease) to cut dna at specific sites. Proteins that acatlyse reactions (excpetion is a small group of catalytic rna molecules) 6 classes: oxidoreductases, transferases, lyases, isomerases, ligases, hydrolases (restriction enzymes are this). Enzymes have a high degree of specificity for substrate and act reversibly via active sites. Require specific conditions: cofactors/coenzymes for some enzymes, temperature, ph.