Extended Data Fig. 8: Cloning scheme of the CRISPR/Cas9 plasmid using a Type IIS restriction enzyme.
From: Intron-encoded cistronic transcripts for minimally invasive monitoring of coding and non-coding RNAs
The empty CRISPR-Cas9–i53 cloning vector is digested with the type IIS restriction enzyme BbsI (isoschizomer: BpiI). The backbone fragment is used for subsequent ligation with a spacer fragment derived from the dimerization of two deoxyoligonucleotides. The ligation product is then used for transformation of competent E.coli.
