Ffpe dna extraction kit

Specimens were preserved in formalin and were transferred for pathological diagnosis and genomic testing. The pathological diagnosis was evaluated after hematoxylin & eosin and immunohistochemistry staining. The DNA/RNA was extracted from formalin-fixed, paraffin-embedded specimens via an FFPE DNA Extraction Kit (Amoy Diagnostics). The genomic testing of all specimens was performed in an AmoyDx Multi-Gene Mutations Detection Kit (Amoy Diagnostics), which harbored DNA-based mutation and RNA-based fusion detection real-time PCR assays, and can detect the mutational status of EGFR, KRAS, BRAF, NRAS, HER2, PIK3CA, ALK, ROS1, MET, and RET simultaneously [22 (link)].

EGFR mutation detection was conducted using the Amplification‐Refractory Mutation System. FFPE DNA extraction kit (Amoy Diagnostics, Xiamen, China) was used to extract tumor DNA. EGFR mutation was detected by EGFR 29 Mutations Detection Kit (Amoy Diagnostics, Xiamen, China) using 80 ng DNA. The procedures were followed‐up as described in the protocol. PCR was performed on a Stratagene Mx3000P cycler (Agilent, Santa Clara, CA, USA) using the following program: Five minutes at 95°C (one cycle); 25 seconds at 95°C, 20 seconds at 64°C, and 20 seconds at 72°C (15 cycles); 25 seconds at 93°C, 35 seconds at 60°C, and 20 seconds at 72°C (31 cycles). The results were determined as described in the protocol.