SMRT-OTS of human,  benchmarking gRNAs, mulitplex: HEK293 DNA
A  SMRT-OTS library was constructed with  a modified version of the protocol described by Tsai et al (2017), BioRxiv. Instead of enzymatic  shearing and genomic complexity  reduction, random shearing, to 8 kb, with Megaruptor 2 (Diagenode) was  used. The sample was also  size-selected using the Blue Pippin system (Sage Science) before going into the Cas9 digestion. Guide  RNAs targeting the EMA1, FANCF, RNF2  and VEGFA genes were used in the experiment. Sequencing was performed  on the Sequel system using 3.0  sequencing and binding chemistry and a 10 hr movie time.