L5 Handout - Lab 5 - GMB I

L5 Handout - Lab 5 - GMB I - Bio 1A Lab Lecture Outline 5...

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Bio 1A Lab Lecture Outline # 5 GMB I Sept. 29 th , 2008 I. Review photosynthesis – controls, measurements of photosynthesis. II. Isolation of Plasmid DNA (mini-prep) A. Pellet, resuspend, lysis (buffer 2 –don't wait), neutralize, pellet, supernantant to column. B. An overview of the functions of each step. III. Restriction digestions. A. Remember the specificity and how it cuts BOTH strands. B. You need to add buffer prior to the enzyme (2X RB)! C. How big, on average, will the pieces be? Making predictions! IV. DNA sequencing A. Key concepts: population of molecules, 5' & 3 & DNA pol, dNTP vs. ddNTP electrophoresis B. Application. – reading the gel! V. Complementation A. 1N strains defective in histidine biosynthesis were mated to create 2N strains. B. Can they now make histidine? Replica plate onto SD plate lacks histidine. C. Compare 2N and 1N. If growth they complement. If no growth they don't complement. VI. Human Life Cycle (7 th & 8 th eds. 13.5, 8 th ) A. Mitosis – when & where B. Meiosis – when & where C. Fertilization – when & where D.
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