RNA-Seq of human bronchial smooth muscle cells: LRP1 knockdown 48 hours
Total RNA was extracted and assessed for quality (Nanodrop, gel electrophoresis, RIN via Agilent 2100). Poly-A mRNA was isolated using Oligo(dT) beads, fragmented to ~300 bp, and reverse transcribed into cDNA with random hexamers. The cDNA was end-repaired, "A" bases were added to the 3' ends, and Y-shaped adapters were ligated. The library was enriched by PCR, gel purified, and quantified using Picogreen. Sequencing was performed on the Illumina NovaSeq 6000 platform (2150 bp, paired-end).