URL: http://www.addgene.org/87424
Proper Citation: RRID:Addgene_87424
Insert Name: DSP Homology Arms with linker-mEGFP
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:28814507
Vector Backbone Description: Vector Backbone:pUC57; Vector Types:Mammalian Expression, CRISPR, Donor Template; Bacterial Resistance:Ampicillin
Comments: This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the C-terminus of human DSP in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: HDPPVAT. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ . Please visit https://www.biorxiv.org/content/early/2017/03/31/123042 for bioRxiv preprint.
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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