Patient screening
tumor
TUMOR
Total RNA was isolated from FFPE tumor blocks (starting from 10 cuts of 10 µm), using the Recover All Total Nucleic Acid Isolation® kit (Ref: AM1975; Ambion, Thermo Fisher Scientific, Waltham, MA, USA), following manufacturers’ instructions. The cDNA synthesis was performed using 200 ng of RNA, random primers, a dNTP mix, and MultiScribe Reverse Transcriptase in a total volume of 25 μL (Ref: 4368814; High Capacity cDNA Reverse Transcription Kit; Applied Biosystems, Waltham, MA, USA). The qPCR reactions were performed in 384-well plates with the TaqMan Gene Expression Assays (Applied Biosystems). To quantify gene expression the following TaqManThe mRNA expression levels of both 