Description
Quantity: 10 Reactions
Unit Size: Each
• A rapid solution to drive high-efficiency gene tagging (up to 100% edited cells) with transfection-ready donor DNA for genome editing experiments
• Kit provides everything needed to produce a high-quality DNA donor to tag the gene with a selection marker and BFP
• Simply provide two locus-specific primers to add homology arms, PCR-amplify, and purify for a transfection-ready donor in hours, not days
• One-step PCR amplification to generate donor templates – no cloning required
• Enrich up to 100% BFP-tagged cellsEach kit is designed to enable users to:
• Tag the carboxy- (C-) or amino- (N-) terminus of the gene of interest
• 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: N-terminus/puromycin resistance, N-terminus/blasticidin resistance, 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 beta-actin (ACTB)Intuitive software to rapidly design gene-specific reagentsTarget-specific gene editing reagents are required for proper knock-in of your 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 the 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
