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Fig. 3 | Orphanet Journal of Rare Diseases

Fig. 3

From: Genotype-phenotype correlation and description of two novel mutations in Iranian patients with glycogen storage disease 1b (GSD1b)

Fig. 3

Long-range PCR and sequencing showed the full-gene deletion of SLC37A4 in the siblings (P5–1 and P5–2) with GSD1b. a the gene transcript image (was taken from Genome Data Viewer in NCBI) and the orientation of the designed primers across upstream and downstream breakpoint. The black arrows indicate the position of primers used in Long-range PCR. The length of the genomic segment for each set of primers (F1&R1 and F1&R2) was shown. b Sanger sequencing result of the breakpoint site and flanking region. Two boxes above the sequencing result indicate the sequences across upstream and downstream breakpoint. In Sanger sequencing result, the blue arrow shows the breakpoint and a 6712 bp sequence deletion on chr11 of the human reference genome (GRCh37). c Gel electrophoresis of the PCR product. i) Results of the first long-range PCR (PCR with F1 and R1 primers) are presented in the left which shows this segment in the siblings, parents and control samples. Lane 1 contains a 10 kb ladder, lane 2 and 3 contains the products of deleted allele, lane 3 and 4 contains the products of both the deleted allele and the wide type. Lane 5 contains the wide-type allele. ii) Results of the second long-range PCR (PCR with F1 and R2) are presented in the right which shows this segment in two siblings, parents and control samples. Lane 1 contains a 10 kb ladder, lane 2 contains NTC, Lane 3, 4 and 5 contains a 2724 bp fragment without deletion and lane 6 and 7 contains no amplification. All lanes (except lane 2) include a ~ 700 bp internal control (Exon 5 of G6PC gene). The products of deleted allele, (1564 bp); the products of wide-type allele, (8276 bp); M, mother; F, father; CT, control sample

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