Nano-OTS of human,  benchmarking gRNAs, multiplex: HEK293 DNA
A  Nano-OTS libary was constructed using  a modified version of the protocol described by Gilpatrick et al. (2020), Nature  Biotechnology. Instead of going in with  intact genomic DNA in the library construction, the DNA was sheared to  20 kb using the Megaruptor 2  (Diagenode) and size-selected prior to library construction. In addition, only one guide  RNA per target was used. Guide RNAs  used in this experiment were designed to target the EMA1, FANCF, RNF2 and VEGFA  genes. Sequencing was performed on the MinION system using one  R9.4.1 flowcell.