title: GSM8793891: EMX1 on- and off-targets; synthetic construct; OTHER
library_name: GSM8793891
library_construction_protocol: Chemically synthesized The synthetic oligonucleotide-based sequencing library was amplified and converted into double-stranded DNA. The PCR reaction was purified using Ampure beads, followed by in vitro cleavage with SpCas9 and Proteinase K treatment. The resulting product was purified with Ampure beads and subjected to end extension and adaptor ligation. The adaptor-ligated product was size-selected for fragments between 100-200 bp using the BluePippin platform. After size selection, the product was purified, amplified, and appropriately diluted. The diluted product was then used for standard barcoding PCR. The final product was sequenced on a MiSeq system following Illumina's standard protocol.