Transformation Lab – A “Thought Experiment” to answer Data Analysis Q#8 on p. 164 Debbie’s Group completes a transformation of E. coli bacteria with a recombinant DNA mixture (RM DNA) that they hope will allow them to clone the Green Fluorescent Protein (GFP) gene (i.e. produce pGLO). After the transformation procedure, they plate some of their E. coli + RM DNA cells on a LB/amp plate and some cells on a LB/amp/ara plate. The group knows that these two plates should both produce transformants, colonies with either pBAD18 or pGLO. They can distinguish which colonies contain pBAD18 from those that contain pGLO on the LB/amp/ara plate because the presence of arabinose causes the expression of GFP in cells with pGLO. Ultimately those cells will fluoresce green under UV light. They wonder, “ How can we screen the colonies from our LB/amp plate to determine which contain the nonrecombinants pBAD18 and which contain the recombinant pGLO? ” They discover that they can duplicate each colony from their transformation onto different media in
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This note was uploaded on 05/22/2011 for the course BIO 205 taught by Professor O'neal during the Spring '08 term at SUNY Stony Brook.