Peripheral blood samples were harvested at diagnosis from BC or OC patients through a vacutainer syringe containing EDTA. Genomic DNA was isolated using the DNeasy® Blood Kit (QIAGEN, Hilden, Germany). After the extraction phase, DNA has been quantified by Qubit®3.0 fluorometer (Thermofisher Scientific, Waltham, MA, USA) and its quality has been assessed through the use of 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
The genetic analysis for BRCA1/2 was performed as previously described (4 (link), 26 (link)).
Sequencing analysis was performed using Ion 520 Chip (Thermofisher Scientific, Waltham, MA, USA) and Ion Torrent S5 (Thermofisher Scientific, Waltham, MA, USA) NGS platform. Obtained data were analyzed using both Amplicon Suite (SmartSeq s.r.l.) and Ion Reporter Software v.5.12 (Thermofisher Scientific, Waltham, MA, USA). NGS data analysis was performed with the standardization of sequencing coverage depth in order to minimize the probability of false positive and negative results in clinical practice, considering a minimum coverage of 500× to each sample.
The genetic analysis for BRCA1/2 was performed as previously described (4 (link), 26 (link)).
Sequencing analysis was performed using Ion 520 Chip (Thermofisher Scientific, Waltham, MA, USA) and Ion Torrent S5 (Thermofisher Scientific, Waltham, MA, USA) NGS platform. Obtained data were analyzed using both Amplicon Suite (SmartSeq s.r.l.) and Ion Reporter Software v.5.12 (Thermofisher Scientific, Waltham, MA, USA). NGS data analysis was performed with the standardization of sequencing coverage depth in order to minimize the probability of false positive and negative results in clinical practice, considering a minimum coverage of 500× to each sample.
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