Plasmid Cloning: Restriction, Ligation & Transformation

This simulator walks through the core genetic-engineering workflow used to insert a gene of interest into a bacterial plasmid: cut both the vector and the insert with a restriction enzyme, ligate them together with T4 DNA ligase, then heat-shock transform the mixture into competent E. coli and select surviving colonies on an antibiotic plate. Choosing mismatched enzymes for vector and insert leaves incompatible sticky ends that ligase cannot join; the insert:vector molar ratio slider controls what fraction of successfully ligated plasmids are recombinant (carrying the insert) versus self-ligated vector; and the heat-shock duration slider controls transformation efficiency along the real bell-shaped curve — too short and the membrane pores never open, too long and the bacteria die. Press Plate Transformation to spread the resulting mixture onto a 3D petri dish and watch green (recombinant) and white (vector-only) colonies grow in the proportions your choices predict.