...be worn at all times. Use plastic pipettes in place of glass pipettes or needles. Liquid waste should be...vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector chosen by the TRC for expression...M0202S T4 DNA ligase buffer NEB #B0202S DH5 alpha competent cells Invitrogen #18258-012 Qiaquick gel extraction...hours. Transform 2 μL of ligation mix into 25 μL competent DH5 alpha cells, following manufacturer’s protocol...OPTI-MEM In a polypropylene tube, add OPTI-MEM first. Pipette FuGENE® directly into the OPTI-MEM – do not allow...tube from step c for a total volume of 100 μL. Pipette master mix directly into the liquid and not onto...cells. Swirl to disperse mixture evenly. Do not pipette or swirl too vigorously, as you do not want to ...