Learning Module - Experiment 4B

Directional cloning of the topAcysB fragment into pTrc99A


We want to subclone the topAcysB insert from lambdaZAPII/topAcysB into the expression vector pTrc99A


The cloning strategy is as follows:

cloning steps


SacI and XbaI have different recognition sites, and leave incompatable ends

overhanging sequences


The insert can only go into the vector in one direction

directional cloning


We will analyze the success of this cloning experiment by using agarose gel electrophoresis in Experiment 5B