Genomic Deletions in Mammalian Cell Lines
Type
Collection
...usage of the pSpCas9(BB) plasmid (pX330) (Addgene plasmid ID 42230). This plasmid allows for the simultaneous...and “AAAC” before the guide’s reverse complement for cloning into the pX330 vector using BbsI restriction...CRISPR/Cas9 Clones for Deletions and Clone Selection For suspension cells, transfer all clones to a single...deletion clones with a proofreading polymerase and clone the amplicons ( e.g., with a PCR cloning kit) into... into a plasmid vector. Transform the plasmid into DH5α E. coli cells and plate onto LB agar plates with...cellular delivery of a pair of chimeric single guide RNAs (sgRNAs) to create two double strand breaks (DSBs... multiple sgRNAs may be expressed from a single plasmid. Add “CACC” before the 20-mer guide sequence and...