2.6. Total RNA extraction and quantitative real-time PCR
PRIMARY TUMOR, METASTASIS, METASTATIC TUMOR
To explore the roles of the miRNAs in CRC metastasis, the expression of the miRNAs was compared between SW480 and SW620 cells, which were isolated from primary tumor and metastatic tumor in abdomen of single CRC patient, respectively. Total RNA was isolated from the cell lines using Total RNA Extractor (No. B511311; Sangon Biotech, Shanghai, China). Reverse transcription reactions were performed using miRNA First Strand cDNA Synthesis Tailing Reaction Kit (No. B532451; Sangon Biotech). The quantitative real-time PCR experiment was performed using a MicroRNAs qPCR Kit (SYBR Green Method) (no. B532461; Sangon Biotech) on an CFX96 Real-Time PCR Detection System (Bio-Rad).