1：Experimental Design and Method Selection:

The study used a double-hybridization approach where mRNA is immobilized via hybridization to DNA capture probes on a microarray and labeled with Cy5-conjugated DNA probes. This avoids enzymatic steps like reverse transcription and PCR.

2：Sample Selection and Data Sources:

Total RNA was isolated from Panc-1 human pancreas carcinoma cells using an RNeasy Mini Kit.

3：List of Experimental Equipment and Materials:

Key equipment included a Microgrid II contact-printer, Axiovert 200 microscope, and various buffers and reagents from suppliers like Sigma-Aldrich and Qiagen. Materials included epoxy-functionalized substrates, amino-modified DNA probes, and Cy5-labeled probes.

4：Experimental Procedures and Operational Workflow:

Microarrays were fabricated by spotting capture probes, followed by hybridization with RNA and label probes under controlled conditions (e.g., denaturation at 70°C, hybridization at 40°C overnight). Fluorescence microscopy was used for detection.

5：Data Analysis Methods:

Data were analyzed using MATLAB for spot brightness and SigmaPlot for statistical analysis, including one-way ANOVA.