Genomic Deletions in Mammalian Cell Lines
Type
Collection
...oligos 1:10 in ddH 2 O ( e.g., 1.0 μl annealed oligos + 9.0 μl ddH 2 O to yield a concentration of 1 μM)....35 cycles of (95 °C for 30 sec, 60 °C for 1 min, 72 °C for 1 min), and 72 °C for 10 min. Optimize PCR ...the page. CRISPR Design Design sgRNAs manually or using freely available online tools 1 . Use these tools...at the genomic recognition site. NOTE: Figure 1 describes possible deletion strategies for genes and non-coding...Use the example guides for the intended deletion of Pim1 in mouse ( Mus musculus ; Table 1 , Figure 2A )...Deletions in Mammalian Cell Lines via CRISPR/Cas9. Bauer DE, Canver MC, Orkin SH. J. Vis. Exp. (95), e52118, ...complement sequences of the Pim1 sgRNA from Table 1 are found in Table 2 . Obtain 24- or 25-mer oligos...