... your gene of interest, in frame with the start codon or tag sequences (where appropriate). The primer...sterile water (instead of TE buffer) to ensure optimal salt concentrations in subsequent reactions. Step...

...Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal 21-mer targets in your gene B.2 Order oligos compatible... F.1 Recommended materials F.2 Determining the optimal puromycin concentration F.3 Protocol for lentiviral...Designing shRNA Oligos for pLKO.1 B.1 Determining the Optimal 21-mer Targets in your Gene Selection of suitable...silencing: Starting at 25nt downstream of the start codon (ATG), search for 21nt sequences that match the ...plasmid to obtain the ratio that gives you the optimal viral production. d. Create a master mix of FuGENE...located in the “Appendix”. F.2. Determining the Optimal Puromycin Concentration Each cell line responds...Addgene strongly recommends that you determine the optimal puromycin concentration for your cell line before...