Bio1AL_Sum09_lab2_lecture

Bio1AL_Sum09_lab2_lecture - Bio 1AL Lab Lecture Outline # 3...

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Bio 1AL Lab Lecture Outline # 3 June 29 th , 2009 I. Constructing recombinant DNA. A. Overview- Isolate, Restriction Digest, Ligate, Transform, Screen! ( I R eally D o L ike T his S tuff. ) B. DNA structure – ds, anti-parallel, base pairing rules: A with T, C with G, bond strengths C. Restriction enzymes – specificity, cleave phosphodiester bond, overhang, sizes of pieces. D. Ligation reaction – annealing, form the phosphodiester bond. i) typically 3 classes of DNA: ii) how to determine which type? Use of selectable genes in the vector. iii) you are using purified λ DNA as the "insert" E. Transformation- uptake and expression of DNA in a host cell i) Host cell = Escherichia coli ( E. coli ) ii) Growth & Division a) needs energy source, carbon source, elements, = LB b) LB plates – LB with agar, one bacterium to a colony c) selectable genes in the vector – eg. ampicillin affects cell wall synthesis iii) Transformation is a rare event. How do you screen for this? for various classes of
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This note was uploaded on 07/04/2009 for the course BIO 1al taught by Professor Pederson during the Summer '08 term at Berkeley.

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Bio1AL_Sum09_lab2_lecture - Bio 1AL Lab Lecture Outline # 3...

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