Fig. 1.

Generation of MYH6-mScarlet iPSC reporter line. A Schematic illustration depicting the MYH6-mscarlet reporter design. B The target-specific PCR designed for screening the reporter integration that spans outside the 5′ homology arm up to the 2A-mScarlet region gives an amplicon of 1.2 kb. The parallel gel image shows the positive amplification of target PCR in the reporter clone D-103. C PCR strategy to identify the homozygosity and Cre removal of puromycin selection cassette. The primers spanning the integration region were used to amplify the genomic DNA from the knock-in clones. Homozygous clone (HM) with the modified locus in both alleles produces only one amplicon of 3.5 kb that comprises the knock-in cassette, whereas the heterozygous clone (HT) produces two amplicons from the modified and wild-type (WT) parental locus of 3.5 kb and 0.8 kb respectively (left panel gel image). The removal of puromycin selection cassette reduces the amplicon length of the modified locus to 1.6 kb as shown in the right panel gel image