pX330-U6-Chimeric_BB-CBh-hSpCas9

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

Construction of Single-Guide RNA Expression Vectors for the CRISPR/Cas9 System
The pX330 plasmid, a bicistronic expression vector containing the cas9 gene and BbsI cleavage sites, was purchased from Addgene (Cambridge, MA, U.S.A.). Single-guide RNAs (sgRNAs) specific for the human OCLN gene were designed based on a published report,18) and sgRNAs with BbsI overhangs were cloned into the pX330 vector. Oligonucleotides used to construct sgRNA expression vectors targeting the OCLN gene were as follows: the site A set, 5′-CAC CGA CAG GAT CCG AAT CAC TCC-3′ and 5′-AAA CGG AGT GAT TCG GAT CCT GTC-3′; and the site B set, 5′-CAC CGG CTT TGG TAG CTA CGG AAG-3′ and 5′-AAA CCT TCC GTA GCT ACC AAA GCC-3′. Constructed vectors were named pX330-site A and pX330-site B, respectively.

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Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing CRISPR Human - Deletion OCLN using pX330-U6-Chimeric_BB-CBh-hSpCas9 from Addgene.

Manufacturer protocol

Download the product protocol from Addgene for pX330-U6-Chimeric_BB-CBh-hSpCas9 below.

Download manufacturer protocol

Videos

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