TrueTag™ Knockout Enrichment Donor DNA Kit (Invitrogen™)

The Invitrogen TrueTag Knockout Enrichment Donor DNA Kit provides 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.

Benefits of the TrueTag Knockout Enrichment Donor DNA Kit include:
• 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 cells

The TrueTag Knockout Enrichment Donor DNA Kit provides everything you need 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. Each kit is designed to enable you 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 enrichment

Included 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 human HPRT1

TrueDesign Genome Editor software to rapidly design gene-specific reagents
Target-specific gene editing reagents are required for proper knock-in of your fluorescent tag. Identify your target gene transcript of interest and the TrueDesign Genome Editor web-based software does the work for you:
1. Displays your target gene transcript
2. Designates your insertion site
3. Designs and enables easy ordering of all required components not included in the TrueTag 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 Donor

Also 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