GSM9896425: MDA-MB-231 siNC biological replicate 2; Homo sapiens; RNA-Seq
Total RNA was extracted using TRIZOL (Ambion, 15596-018), further purified by phenol-chloroform extraction, and treated with RQ1 DNase (Promega, M6101) to remove DNA. RNA purity and quantity were assessed spectrophotometrically, and integrity was verified by 1% agarose gel electrophoresis. For each sample, 1 microgram total RNA was treated with RQ1 DNase. Poly(A)+ mRNA was captured using VAHTS mRNA Capture Beads (Vazyme, N401). Directional libraries were prepared using the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina (Vazyme, NR605) and VAHTS RNA Multiplex Oligos Set 1 for Illumina (Vazyme, N323). The dUTP-marked second cDNA strand was not amplified, enabling strand-specific sequencing. Libraries were sequenced as 150-nt paired-end reads on an Illumina NovaSeq X Plus system.