Supplementary Material for Vicencio et al., 2019

Figure S1 shows the correlation between the number of dpy-10 edits and successful edits in the target locus. Figure S2 shows the reagents and conditions for the generation of universal PCR product repair templates for Nested CRISPR Step 2. Figure 3 contains the standard composition of injection mixes for Nested CRISPR. Table S1 contains a summary of experiments with megamers. Table S2 lists the sequences of ssODN bridges in experiments coupled with megamers. Table S3 lists the sequences of the GFP and mCherry megamers. Table S4 contains a list of crRNAs used for Nested CRISPR Step 1. Table S5 contains a list of ssODNs used for Nested CRISPR Step 1. Table S6 contains a list of Nested CRISPR universal sequences. Table S7 contains a list of the strains generated in this study. Table S8 contains a list of external primers for genotyping. Video S1 illustrates the pipeline for generating translational reporters; the fluorescent protein (FP) 1-3 fragment is inserted in step 1 and serves as homology regions for the insertion of the longer remaining FP sequence in step 2. Video S2 illustrates the pipeline for generating a knockout in step 1 and the corresponding transcriptional reporter in step 2.