pX330A-1x3

Protocol tips

Upstream tips	Protocol tips	Downstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.

Upstream tips
To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip (20uL or 200uL one), puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate.

Publication protocol

Genome editing using the multiplex CRISPR/Cas9 assembly system
The specific gRNA sequences were selected using the CRISPR design tool (http://crispr.mit.edu/) and cloned into the pX330A-1 × 3 multiplex gRNA assembly vector (Addgene plasmid #58767)50 using the Golden Gate cloning method. Then, the multiplex gRNA cassette was replaced with the lentiviral vector LentiGuide-Puro (Addgene plasmid #52963). Stable expression of Cas9 and gRNAs was established via puromycin or blasticidin resistance using LentiCas9-Blast (Addgene plasmid #529632) following Zhang et al.’s protocol51.

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Reviews

Discussion

Papers

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Manufacturer protocol

Download the product protocol from Addgene for pX330A-1x3 below.

Download manufacturer protocol

Videos

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