Transformation Of Expression Plasmid Constructs

Protocol Overview

Thaw One shot Top10 chemically competent cells on ice completely and then add plasmid construct (500 ng, ideally in less than 5 microliter volume). Mix the contents by flicking the bottom of the tube and then incubate on ice for 30 minutes. Heat shock the cells at 42 degree C for 45 seconds. Immediately place the heat shocked cells on ice for 5 minutes. Then recover these cells in 250 microliter of SOC medium at 37 degree C for 2 hrs. Plate appropriate dilutions of recovered transformation mix onto LB agar with chloramphenicol at 10 microgram/ml. Incubate plates at 37 degree C.