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How do I enrich HDR edited cells without antibiotic selection or a fluorescent reporter?
We recommend optimizing transfection conditions of NHEJ-mediated editing prior to including the donor oligo (Cas9 nuclease + highest functional crRNA:tracrRNA). After efficient NHEJ-mediated editing is achieved we recommend optimizing the amount of the donor template. Using this method, we found 2.5 nM to 10 nM concentrations of the donor oligo were optimal for U2OS cells.