Fig. 2

Advantages of the modified targeted capture-based protocol over traditionally applied multiplex PCR-based NGS of two low quality DNA-samples extracted from FFPE tissues of patients P5 and P10. a In contrast to the modified targeted capture (TP)-based approach, pathogenic BRCA1 variants were missed by the multiplex PCR (MP)-based approach. b DNA concentration of FFPE samples of patients P1 to P13. The FFPE samples of P5 and P10 are marked by a red dot. c Delta ct values of DNA from FFPE samples of patients P1 to P13. The DNA of FFPE samples of P5 and P10 are marked by a red dot.d Zero-covered nucleotides occur only with the multiplex PCR-based protocol. e Low-covered nucleotides (< 500 with the multiplex PCR-based approach and < 30 with the targeted capture-based approach) occur only with the multiplex PCR-based protocol. f Artifacts only occur with the multiplex PCR-based approach. g Polymorphisms were detected via both approaches. In patient 10 two additional polymorphisms were found with the targeted capture-based protocol. h False-positive pathogenic BRCA variants were detected in both patients using the multiplex PCR-based technique