Fig. 2

Molecular analysis of a patient family carrying the novel MSH6 exon 5–6 skipping variant. (A) The names, sequences, positions, and corresponding amplicon sizes of primers used in the RT- and genomic PCR analysis for the MSH6 variant screening. (B) Mutant variant detection by RT- and genomic PCR and agarose gel electrophoresis. The RT- and genomic PCR reactions were performed with primers spanning the skipping exons listed below the gel images and compared between the patient (I-1), normal control (HEK293 cell line), and negative control (NC, distilled water). The asterisk (*) labels the mutant variant amplicon in RT-PCR, while the pound sign (#) indicates the mutant variant amplicon in genomic PCR. (C) The Sanger sequencing results of variant fragments amplified from RT- or genomic PCR. The variant is described in relation to the reference sequence following the HGVS nomenclature (version 20.05). (D) Pedigree and RT-PCR analysis of the patient family with the novel MSH6 variant. In the pedigree, filled symbols represent affected individuals, and the arrow indicates the proband. The RT-PCR gel shows the presence of the variant (indicated by the lower band) in the proband and two of his sons