TrueTag™ Donor DNA Kit, RFP stem (Invitrogen™)

The Invitrogen TrueTag Donor DNA Kit, RFP stem, provides a rapid solution to drive high-efficiency tagging (up to 100% edited cells) of silent or differentially expressed genes with transfection-ready donor DNA.

Benefits of the TrueTag Donor DNA stem kits include:
• One-step PCR amplification to generate donor templates—no cloning required
• Generate transfection-ready DNA donors in hours instead of days
• Save up to three months with enrichment compared to single cell isolation and characterization
• Enrich up to 100% RFP-tagged cells

The TrueTag Donor DNA Kit, RFP stem, provides everything you need to produce a high-quality DNA donor to tag your gene with a selection marker and RFP—simply provide two locus-specific primers to add homology arms and PCR-amplify the donor. Each kit is designed to enable you 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 your cell model
• Easily remove antibiotic resistance markers with Cre/Lox following cell enrichment

Included 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)

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