MCB 2050 Study Guide - Midterm Guide: Huntingtin, Forensic Identification, Heat Shock Protein
14 Feb 2017
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Category 1: basic techniques used to clone genes. Cystic fibrosis: 1/2000 newborns, autosomal recessive, mucus accumulation, 32 year life expectancy. Screen: step 4: amplify grow in broth, lyse, isolate, analyze. Cut at specific sequences (restriction sites) and break phosphodiester bonds) can be cohesive or blunt. Discovery: 1970 smith and nathans: re protect bacteria from viruses. Protect bacterial genome from re by methylation of nucleotides within recognized sequence. Nomenclature: named according to bacterial strain (escherichia coli strain ry13 restriction enzyme number i) Recognize palindromes makes staggered or blunt. Complimentary strands made by re can be ligated together using dna ligase recombinant. Amplification of recombinant dna molecules in cloning vectors: Cloning vectors used to isolate and amplify dna sequences. 3 parts: origin of replication for amplification, dominant selectable marker (antibiotic resistance, unique re cleavage site outside of origin and resistance gene (often mcs all. Re sites in mcs are unique in plasmid)
