Spacer_Wt_set2
Expanded CRISPR arrays corresponding to the expression of Wt Cas1-2 was extracted from the agarose gel (QIAquick Gel Extraction Kit, Qiagen). Approximately 200 ng of the PCR product was further purified using HighPrep magnetic beads (MAGBIO). This purified sample was subjected to DNA end repair and adaptor ligation (5-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3 and Index Adapter:5-GATCGGAAGAGCACACGTCTGAACTCCAGTCAC [INDEX] ATCTCGTATGCCGTCTTCTGCTTG-3) using Illumina-compatible NEXTflex Rapid DNA sequencing kit (BIOO Scientific, Austin, Texas, U.S.A.). Subsequently, the ligated DNA products were purified with HighPrep magnetic beads and further enrichment was achieved by 8 cycles of PCR with Illumina-compatible primers (NEXTFlex DNA sequencing kit). These amplicons were subjected to an additional step of purification with HighPrep magnetic beads and were sequenced on a Miseq 300 paired end platform.