ELANE Cas9WT
Library preparation was performed using a two-step PCR method. For the first PCR reaction, a pair of target-specific primers was designed to amplify the 150bp area surrounding the cutting site. Each target primer additionally includes an extension at the 5 end: for forward primers, this contains the Illumina Read 1 primer sequence (see below, nucleotides in bold) and an 8bp Unique Molecular Identifier (UMI, nucleotides underlined), and for the reverse primers, this contains the Illumina Read 2 primer sequence only (see below, nucleotides in bold):RNF2 fwd 5-3: ACA CTC TTT CCC TAC ACG ACG CTC TTC CGA TCT NNN NNN NNG ACA AAC GGA ACT CAA CCA TRNF2 rev 5-3: GTG ACT GGA GTT CAG ACG TGT GCT CTT CCG ATC TTG TTC TAT TTA AGT TTT CAT GTT CTELANE fwd 5-3: ACA CTC TTT CCC TAC ACG ACG CTC TTC CGA TCT NNN NNN NNC TCC CCG GCA GAA ACG TCELANE rev 5-3: GTG ACT GGA GTT CAG ACG TGT GCT CTT CCG ATC TGA GAA TCA CGA TGT CGT TGA GC