GSM9410081: WM4011-2_Baseline_6125; Homo sapiens; RNA-Seq
Total RNA was extracted from snap-frozen PDX tumor tissues using standard RNA extraction protocols. RNA quantity and quality were assessed prior to library preparation. RNA sequencing libraries were prepared at the Broad Institute following the Broad Stranded RNA-Seq protocol. Total RNA was quantified using the Quant-iT™ RiboGreen® RNA Assay Kit and normalized prior to library preparation. ERCC spike-in controls were added to each sample. Libraries were constructed using an automated version of the Illumina TruSeq™ Stranded mRNA Sample Preparation Kit, which preserves transcript strand orientation. Poly(A)+ mRNA was enriched using oligo-dT beads, followed by fragmentation, cDNA synthesis, adapter ligation, and PCR enrichment. Libraries were quantified, pooled, and sequenced on an Illumina NovaSeq platform using paired-end sequencing.