URL: http://www.addgene.org/118055
Proper Citation: RRID:Addgene_118055
Bacterial Resistance: Ampicillin
Vector Backbone Description: Vector Backbone:pSpCas9(BB)-2A-Puro (PX459) V2.0 (Addgene#62988); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: pSpCas9(BB)-2A-Puro (PX459) V2.0 was digested with EcoRI to remove the T2A-Puro cassette, and a P2A-Blasticidin cassette was amplified by PCR, digested with EcoRI, and ligated. The BbsI site (GAAGAC) in the blasticidin-resistance gene was mutated to GAGGAC with no change in amino acids. gRNA inserts can be sequenced using pLKO.1 5’ primer, 5’-GACTATCATATGCTTACCGT-3’.
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Source: Addgene (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org