Description
Quantity: 10 Reactions
Unit Size: Each
• A rapid solution to drive high-efficiency gene knockout and enrichment (up to 100% edited cells) with transfection-ready donor DNA for genome editing experiments
• Kit provides everything needed to achieve functional gene knockout and enrich the edited cells
• The donor produced will insert 3X STOP codons, a selection marker, and a fluorescent tag
• Simply provide two locus-specific primers to add homology arms and PCR-amplify the donor
• Easily design and order all necessary components to generate a knockout cell line
• Save up to 3 months with enrichment compared to single cell isolation and characterization
• Leverage a selection marker to obtain up to 100% edited cellsEach kit is designed to enable users to:
• Insert 3X STOP codons to knock out gene function
• Tag each allele with a different fluorophore (GFP or RFP) to verify bi-allelic knockout
• Utilize puromycin and blasticidin selection markers for bi-allelic knockout confirmation
• Easily remove antibiotic resistance markers with Cre/Lox following cell enrichmentIncluded in the kit:
• Four linear donor templates: GFP/puromycin resistance, GFP/blasticidin resistance, RFP/puromycin resistance, RFP/blasticidin resistance
• Phusion Flash High-Fidelity PCR Master Mix
• PCR cleanup columns and buffers
• Positive control primers for knockout of humanHPRT1Intuitive software to rapidly design gene-specific reagentsTarget-specific gene editing reagents are required for proper insertion of the knockout enrichment DNA donor. The Invitrogen TrueDesign Genome Editor web-based software does the design work for you:
• Identifies target gene transcript
• Selects a pre-designed or custom guide RNA
• Generates primers and enables easy ordering of all necessary componentsRequired reagents available via TrueDesign software (not included in kit):
• CRISPR guide RNA or TALENs designed for high-efficiency cleavage
• 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
