xgen exome hyb panel v2  (Integrated DNA Technologies)

Journal: Frontiers in Medicine

Article Title: Case Report: Beyond type 1 diabetes: a case of delayed MODY1 diagnosis and successful transition to sulfonylurea therapy

doi: 10.3389/fmed.2025.1590935

Figure Lengend Snippet: The novel HNF4A variant affects the splicing of the gene. RT-PCR products obtained from Hek-293 cells transiently transfected with the different pSPL3 constructs. A band of 263 bp was detectable both in cells transfected with the empty vector (EV) and with the construct carrying the third HNF4A exon 3 and flanking intronic sequences with the c.391G>A variant (V). This is due to the skipping of the HNF4A exon induced by the presence of the substitution with combination of the two artificial exons as observed in cells expressing the empty vector. A band of 358 bp is detectable in cells expressing the wild-type minigene construct (WT) and corresponding to the correct splicing combining the exon 3 of the HNF4A gene with those provided by the splicing vector, as schematically represented in the middle panel. The splicing events were also checked by Sanger sequencing of the RT-PCR products as shown by the chromatograms reported in the right panel. 1kb and 100bp, molecular weight markers; -, PCR reaction negative control; SA and SD6, artificial exons provided by the pSPL3 vector.

Article Snippet: 50–100 ng of genomic DNA was used to sequencing selection of coding genomic regions and flanking intronic sequences using IDT xGen Exome Research Panel v2 enrichment kit (34 Mb, 19,433 genes) and Illumina technology (PE 2X150) on the Illumina platform Novaseq6000.

Techniques: Variant Assay, Reverse Transcription Polymerase Chain Reaction, Transfection, Construct, Plasmid Preparation, Expressing, Sequencing, Molecular Weight, Negative Control