Targeted sequencing of post-selection 5xNNS PID library: CAG[GCC] dual reporter and 2mM 3-AT, replicate 1
A single PCR step (15 cycles) generated dual-indexed amplicons spanning the 5xNNS-randomized region of an omega-dSpCas9 PID library used in B1H selection experiments; forward and reverse primers provided the appropriate Illumina indices and adapters needed for direct sequencing on a NextSeq 500. Amplicons were size-selected by gel purification before pooling for a multiplexed, paired-end sequencing run (75 x 75) that captured both strands of the NNS-randomized region entirely.