title: Design 1_Golden Gate_BsaI at Amplicon stage_SR3
design: First, gRNA oligonucleotides were annealed to generate fragments containing the gRNA spacers flanked by BsaI recognition sites. These fragments were then digested with restriction enzymes and ligated to vectors containing a promoter and terminator for gRNA expression. The PCR amplicons of the ligation products during plasmid construction (~400 bp)  was prepared. Nanopore sequencing of PCR amplicons were prepared using SQK-RBK110.96.