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Survival during cardiac directed differentiation does not depend on initial human induced pluripotent stem cell differences
Version 2 2022-02-05, 19:30
Version 1 2022-02-05, 18:00
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posted on 2022-02-05, 19:30 authored by Connie Jiang, Rajan Jain, Arjun RajArjun Raj| Targeted sequencing of barcodes from genomic DNA harvested from hiPS cell-derived cardiomyocytes and hiPS cells, of which only a subset had barcodes. hiPS cells were lentivirally transduced at different MOI with barcode libraries consisting of random 100-mers in the 3’ untranslated region of a green fluorescent protein (GFP) transgene after which they were permitted to divide approximately three times before being split and seeded for two parallel differentiations. On day 15 of differentiation (17 days after seeding), we isolated genomic DNA from cells (after fluorescent activated cell sorting (FACS) gated on GFP positivity for one condition), then performed targeted amplification of the barcode vector for sequencing. See Jiang et al. citation for additional details. |