Neoplastic cellularity was assessed by microscopic examination and, when below 50%, enriched by manually microdissecting four consecutive 10 µm thick sections. All samples were microdissected excluding the five SPT, HCC1 to HCC4, and ICC1.
Genomic DNA from frozen or FFPE tissues was extracted using the QiAamp DNA Mini Kit or QIAamp DNA FFPE Tissue Kit (Qiagen), respectively. Purified DNA was quantified and its quality assessed using Qubit (Invitrogen Life Technologies) and NanoDrop (Invitrogen Life Technologies) platforms [10] (link). DNA suitability for PCR downstream applications was further evaluated through BIOMED 2 PCR multiplex protocol and the PCR products were evaluated by DNA 1000 Assay (Invitrogen Life Technologies) on the Agilent 2100 Bioanalyzer on-chip electrophoresis (Agilent Technologies) [11] (link).
Genomic DNA from frozen or FFPE tissues was extracted using the QiAamp DNA Mini Kit or QIAamp DNA FFPE Tissue Kit (Qiagen), respectively. Purified DNA was quantified and its quality assessed using Qubit (Invitrogen Life Technologies) and NanoDrop (Invitrogen Life Technologies) platforms [10] (link). DNA suitability for PCR downstream applications was further evaluated through BIOMED 2 PCR multiplex protocol and the PCR products were evaluated by DNA 1000 Assay (Invitrogen Life Technologies) on the Agilent 2100 Bioanalyzer on-chip electrophoresis (Agilent Technologies) [11] (link).
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