1. Student work stations for pGLO lab CaCl • Each student group will need four tubes. Label all tubes on the lid/top • Label Tube 1 “LB” (for Luria-BertaniaBroth) • fill with 1mL of LB • Label Tube 2 “CaCl” (for Calcium Chloride Sol’n) • Fill with 1mL of CaCl • Label Tube 3 “+ DNA” • this tube gets the plasmid and is the “experimental chamber” • Label Tube 4 “- DNA” • This tube doesn’t receive plasmid and is the “control chamber” • Place all four tubes in that “floatie” • but if you have microfuge tube racks, use them. • You can choose your own “color coding” system • View from above: LB +DNA -DNA

2. Schematic Diagram of control (-DNA)and experimental (+DNA) tubes 3) 250 ul LB 3) 250 ul LB 2a) one Colony of E. coli 2) one Colony of E. coli 2b) 8-10 ul pGLO plasmid 1) 250ul CaCL 1) 250ul CaCL -DNA (total volume = 500 ul*) +DNA (total volume 510 ul*) *aliquot 100 ul of tube contents onto the appropriate plate and spread. Follow protocol.