Fig. 2: We used capillary electrophoresis to verify the presence of the c.18_28delAGAGAGGCCAA allele. | Human Genome Variation

Fig. 2: We used capillary electrophoresis to verify the presence of the c.18_28delAGAGAGGCCAA allele.

From: High-resolution melting analysis coupled with next-generation sequencing as a simple tool for the identification of a novel somatic BRCA2 variant: a case report

Fig. 2

Although it was difficult to distinguish the results because of this method’s lower resolution, amplification of the somatic mutated DNA gives two PCR products (b): the 218-bp peak of the WT allele and 207-bp peak corresponding to the BRCA2 c.18_28delAGAGAGGCCAA allele compared to the size marker (a). Normalized and shifted melting curves (c) and normalized and temperature-shifted difference plots (d) of the c.18_28delAGAGAGGCCAA allele are shown. Melting profile evaluation of the patient shows a specific melting behavior, as observed in both the normalized and the temperature-shifted and difference plots compared to the FT samples (n = 10) that do not carry this variant. Each experiment is reported in duplicate. The same forward primer used for sequencing was used for PCR-HRMA, while the PCR-HRMA-reverse primer was 5′-TCATTAGGGAGATACATATGGA-3′. The PCR-HRMA primers were designed using Primer3 software (http://bioinfo.ut.ee/primer3) and certified as high molecular-quality products via HPLC purification (Eurofin MWG Operon, Ebersberg, Germany)

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