Sample_GW001_REV_020
MIPs were designed according to previously published protocols. Briefly, the 70 bp flanking the predicted cut site of each sgRNA of all 278 unique sgRNA were chosen as targeting regions, and the bed file with these coordinates was used as an input. Since Trp53 sg4 targets a similar region as the p53 sgRNA within the base vector, the same MIP was used to sequence both of these loci.These coordinates contained overlapping regions which were subsequently merged into 173 unique regions. Each probe contains an extension probe sequence, a ligation probe sequence, and a 7 bp degenerate barcode (NNNNNNN) for PCR duplicate removal. A total of 266 MIP probes were designed covering a total amplicon of 42,478 bp. MIP target size stats: min = 155 bp, max = 190 bp, mean = 159.7 bp, median = 156.0 bp (supplemental table). Each of the mTSG-MIPs were synthesized using standard oligo synthesis (IDT), normalized and pooled.