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URL: http://www.addgene.org/101785

Proper Citation: RRID:Addgene_101785

Insert Name: TUBA1B Homology Arms with mTagRFPT-linker

Organism: Homo sapiens

Bacterial Resistance: Ampicillin

Defining Citation: PMID:

Vector Backbone Description: Backbone Marker:New England BioLabs; Vector Backbone:pUC19; Vector Types:Mammalian Expression, CRISPR, Other, Donor Template; Bacterial Resistance:Ampicillin

Comments: This plasmid has been used with locus-specific CRISPR/Cas9 to add a mTagRFP-T tag to the N-terminus of human TUBA1B in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: GGSGGS. 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/ .

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