...plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal 21-mer targets in your gene B.2 Order...search for “pLKO”“. Back to Top B. Designing shRNA Oligos for pLKO.1 B.1 Determining the Optimal 21-mer...NEB #M0202S T4 DNA ligase buffer NEB #B0202S DH5 alpha competent cells Invitrogen #18258-012 Qiaquick gel...Transform 2 μL of ligation mix into 25 μL competent DH5 alpha cells, following manufacturer’s protocol. Plate ...will alleviate concerns about off-target effects. B.2 Ordering Oligos Compatible with pLKO.1 To generate...insert your sense and antisense sequences from step B.1 into the oligos below. Do not change the ends; these...digestion with EcoRI and AgeI. The oligos from section B contain the shRNA sequence flanked by sequences that...