Description
Quantity: 10 Reactions
Unit Size: Each
• A rapid solution to drive high-efficiency tagging (up to 100% edited cells) of silent or differentially expressed genes with transfection-ready donor DNA
• Kit provides everything needed to produce a high-quality DNA donor to tag gene with a selection marker and GFP Stem
• Simply provide two locus-specific primers to add homology arms and PCR-amplify the donor
• One-step PCR amplification to generate donor templates – no cloning required
• Generate transfection-ready DNA donors in hours instead of days
• Save up to 3 months with enrichment compared to single cell isolation and characterization
• Enrich up to 100% GFP Stem-tagged cellsEach kit is designed to enable users to:
• Enrich tagged cells even when the target protein is not being expressed
• Choose between a fusion or co-expressed fluorescent tag
• Choose from puromycin or blasticidin selection markers – whatever best suits the cell model
• Easily remove antibiotic resistance markers with Cre/Lox following cell enrichmentIncluded in the kit:
• Four linear donor templates: C-terminus/puromycin resistance + 2A, C-terminus/blasticidin resistance + 2A, C-terminus/puromycin resistance, C-terminus/blasticidin resistance
• Phusion Flash High-Fidelity PCR Master Mix
• PCR cleanup columns and buffers
• Positive control primers to tag human glial fibrillary acidic protein (GFAP)Intuitive software to rapidly design gene-specific reagents Target-specific gene editing reagents are required for proper knock-in of the fluorescent tag. The Invitrogen TrueDesign Genome Editor web-based software does the design work for you:
• Identifies target gene transcript
• Designates insertion site
• Designs and enables easy ordering of all necessary componentsRequired reagents available via TrueDesign software (not included in kit):
• CRISPR guide RNA or TALENs designed adjacent to N- or C-terminus
• Oligos to PCR-amplify your donor and add appropriate homology arms
• Sequencing primers for assessment of CRISPR-Cas9 cleavage efficiency
• Verification primers to validate proper insertion of the TrueTag DNA DonorAlso required and available for purchase separately or as part of TrueDesign workflow:
• TrueCut Cas9 Protein v2, TrueCut HiFi Cas9 Protein, or GeneART CRISPR mRNA
• CRISPRMAX Transfection Reagent or MessengerMAX Transfection Reagent
