Extended Data Fig. 5: Confirmation of the ddPCR probe specificity and target SLC30A8 mRNA sequencing.
From: Loss of ZnT8 function protects against diabetes by enhanced insulin secretion
a, R138 (pGEM_CT) and X138 (pGEM_TT) sequences were inserted in pGEM vector and used as template for digital droplet PCR. Original probe configuration confirmed specificity as R138 droplets were only detected by FAM (CT-FAM; channel 1) and X138 droplets were only detected by VIC (TT-VIC; channel 2). In the swapped probe configuration, FAM and VIC probes were swapped and ddPCR was performed using pGEM_CT or pGEM_TT as template. b, Detection of R138 allele (Channel 1) and X138 allele (Channel 2) using cDNA from the heterozygous hiPSC-derived beta-like cells (B1 clone) as a template. In the swapped probe configuration, FAM and VIC probes were swapped and ddPCR was performed using cDNA from hiPSC-derived beta-like cells as template. c, Depicting the unique sequencing reads coverage at p.Arg138* and p.Ala139Ala obtained by SLC30A8 target mRNAs sequencing in edited clones (B1 and A3) and unedited cells (wildtype).
