BIO370Y5 Lecture Notes - Lecture 4: Silent Mutation, Purine, Genentech
Document Summary
January 18th, 2016: purification and identification/ characterization of transformants and inserts. These are the three steps needed to amplify and identify. Detergent (sds) lyses (breaks) the bacteria, and naoh denatures the dna. Potasium acetate/acetic acid returns to neutral ph and percipitates lipids and big proteins. Partially renatured dna is in precipitate while the plasmid dna is in the solution. Centrifugation seperates the dna from the precipitate: early experments in molecular cloning. Chronology of main events relating to development of methdos for constructing and cloning recombinant dnas. 1969-1970 p. berg (1970) and lobban (1969: think about recombinant dna in vitro and using them for cloning and stuff. 1971 d. berg et al (1974: isolate first plasmid dna. 1971-1972 jackson et al. (1972) and lobban and kaiser (lobban 1972; lobban and kaiser 1973: develop terminal transferase tailing, which joins two dnas together, concerns arise about using sv40 dna to make recombinant dna molecule.
