title: iGuideSeq (-) library of 293T cells nucleofected with Let7c1 gRNA
design: gDNA was fragmented to 500 bp average size using a diagenod sonicator, end-repared and ligated with specific adapter. iGuideSeq library was then obtained using 2 round of nested PCR amplification as described in Nobles, C.L., Reddy, S., Salas-McKee, J. et al. iGUIDE: an improved pipeline for analyzing CRISPR cleavage specificity. Genome Biol 20, 14 (2019). https://doi.org/10.1186/s13059-019-1625-3. During the 2nd round of nested PCR, the following barcodes were added : barcode1: GGACTCCT, barcode 2: GTAAGGAGNNWNNWNN