Cell tissue total rna isolation kit v2

The Cell/Tissue Total RNA Isolation Kit V2 (RC112-01, Vazyme, Nanjing, China) was employed to obtain RNA specimens from treated and control cells. The isolated RNA samples were then subjected to mRNA purification and quantification. The mRNA molecules were fragmented using divalent cations and subsequently used for PCR amplification. The resulting PCR products were used to construct libraries for sequencing. Paired-end sequencing was conducted on the system of Illumina HiSeq 4000 (Illumina, San Diego, CA, USA) following the manufacturer’s recommendations. To guarantee the robustness and reproducibility of the results, each experimental group was composed of 3 biological replicates and 3 technical repetitions.
For data analysis, the GO (GO, http://www.geneontology.org/) and Kyoto Encyclopedia of Genes and Genomes (KEGG, http://www.genome.jp/kegg) databases were utilized to identify pathways enriched with statistically significant genes. The analysis aimed to uncover functional insights and molecular pathways associated with the experimental conditions. The technical and methodological aspects of whole transcriptome sequencing were supplied by Majorbio Biotechnology Co., Ltd. (Shanghai, China), ensuring expertise and quality in the sequencing process.

Cell/Tissue Total RNA Isolation Kit V2 (RC112-01, Vazyme, Nanjing, China) was utilized to isolate the Total RNA, based on the manufacturer’s directions. Subsequently, the HiScript III 1st Strand cDNA Synthesis Kit (R312-01, Vazyme, Nanjing, China) was employed to synthesize cDNA. To establish a standard curve and calculate the relative target concentration (qTOWER, Analytik Jena, Jena, Germany), the Sybr Green Master Mix (#11203ES03, Yeasen, Shanghai, China) was utilized to amplify the diluted cDNA samples. The relative expression levels of RNA among different groups were evaluated using the 2^−ΔΔCt method. The sequences of the primer employed in this investigation are illustrated in Table S1.