MCB 2050 Lecture Notes - Lecture 2: Multiple Cloning Site, Dna Ligase, Lac Operon

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Lecture 1 + 2: basic techniques used to clone enzymes. What does each gene do? (want to know what every gene in genome does?) How do we find the genetic mutations that make us sick? (how is it expressed? to what levels? mutations?) What are the normal and mutant gene sequences? (if you locate it, you can clone it and figure out what is normal) To study the function of an individual gene you must be able to isolate it and produce large quantities of it in vitro. Isolate plasmid dna from lysed e. coli cells: analyze the plasmid dna (ex. size, sequence) Dna molecules can be cut at specific sites with restriction enzymes. Answer: to cut dna, you need restriction enzymes, that cut at specific sequences. The bacterial genome is protected from re degradation by methylation of nucleotides within the sequence recognized by the. Dna ligase to create a recombinant dna molecule.

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