Dna microarray scanner g2565b

Total RNA was extracted from individual cell samples using RNA Extraction Reagent (Vazyme, Nanjing, China) and reversely transcribed into cDNA using the HiScript III RT SuperMix for qPCR (+gDNA wiper) (Vazyme), according to the manufacturer’s instructions. The RT-qPCR reactions were performed in triplicate with the ChamQ Universal SYBR qPCR Master Mix (Vazyme) and specific primers (Table S2) in the iQ5™ quantitative PCR detection system (Bio-Rad, Richmond, CA, USA). The data were analyzed using 2^−ΔΔCt method.
Total RNA was also used for analysis of Human OneArray Gene Expression Profiling (Phalanx Biotech, Hsinchu City, China Taiwan) using log2 |FC | ≥ 1 and adjusted P < 0.05 to identify DEGs. After hybridization, the arrays were washed, scanned and the gene expression results were extracted by DNA Microarray Scanner G2565B (Agilent, California, USA), according to the manufacturer’s instructions. Raw fluorescence intensity values were normalized and log-transformed using GeneSpring GX 10 software (Agilent).

Gene microarray analysis was performed by Phalanx Biotech Group (Hsinchu, Taiwan) and included RNA amplification, labeling of probes, hybridization, and data extraction. Briefly, total RNA was extracted from DLD1-PURO cells after three time individual 5-FU treatment as a control group (PURO_H001, PURO_H002, PURO_H003) and DLD1-NGFR cells after three time individual 5-FU treatment as an experimental group (P75_H004, P75_H005, P75_H006) using a Trizol reagent (TaKaRa Bio Inc., Mountain View, CA, USA) according to the manufacturer’s instructions. Gene expression profiling was performed using a Human OneArray v6.1 microarray (Phalanx Biotech Group, Hsinchu, Taiwan) containing 31,741 human genome probes and 938 experimental control probes. After hybridization, the arrays were washed and scanned, and then gene expression results (Figures S1 and S2) were extracted using a DNA Microarray Scanner G2565B (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s instructions. Raw fluorescence intensity values (Figure S3) were normalized and log-transformed using Cluster 3.0 (University of Tokyo, Japan).