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