Figure 2 | Scientific Reports

Figure 2

From: Establishment and characterization of a novel human induced pluripotent stem cell line stably expressing the iRFP720 reporter

Figure 2

Genetic screening. (a) Junction PCRs were performed using locus-specific primers that bind to genomic sequences outside of the homology regions in combination with vector-specific primers. Expected fragment sizes for positive samples: 1196 bp for the left region and 1789 bp for the right region. Three clones tested positive in the screening that contained correctly integrated donor DNA at both ends. C+: positive control hiPSC line containing genome integrated eGFP sequence in the AAVS1 locus, C-: negative control SBAD2 hiPSC line, NTC: no template control. The original uncropped gels are presented in Supplementary Figure S1. (b) Southern blot analysis of the candidate clones (A5-04, A7-09, A7-10) and negative control SBAD2 hiPSC line. gDNA samples were digested with NcoI or ApaI restriction enzymes and tested with AAVS1-LHA or AAVS1-RHA specific probes, respectively. WT: negative control SBAD2 hiPSC line, M: DIG-labeled DNA Molecular Weight Marker VII (Roche), the ladder scale specified by the manufacturer is shown on the right side of the blots. The original uncropped blots are presented in Supplementary Figure S2.

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