Overview: This lab is designed as an extension for the Redesigning Bacteria Transformation with pUC19 plasmid. Students will confirm transformation by PCR amplification of the ampR gene. The results of the PCR reaction will be determined by agarose gel electrophoresis.
Curriculum Connections: bacteria transformation • recombinant DNA • polymerase chain reaction (PCR) • gel electrophoresis
Experimental Timing: From start to finish this lab takes 90-120 minutes. However, there are many good stopping points in this protocol that make it possible to complete the lab in a series of 45-50 minute periods. For additional information, see "Instructor Planning Guide" in the Teacher Materials.
Specialized Equipment Required: p20 micropipettes •p1000 micropipettes (optional) • thermocycler (each student group will set up 3 PCR samples) • gel electrophoresis units with power supplies (each student group will run 4 samples and each gel must also have a lane available for DNA ladder) • UV or blue light source • centrifuge (optional)