title: GSM8975409: Beas-2B-siSPRY2-wsp-4-2; Homo sapiens; RNA-Seq
library_construction_protocol: RNA extraction from lysates harvested in trizol was performed using the purelink RNA kit (Life Technologies). RNA sequencing libraries were prepared according to the Watchmaker mRNA Library Build user guide. Briefly, mRNA from 200ng of total RNA was isolated using polyA, oligo-dT magnetic beads. The isolated mRNA was then subject to enzymatic fragmentation, resulting in ~200 bp fragments. The resulting RNA fragments underwent first and second strand cDNA synthesis. IDT xGen Stubby Adapters were then ligated to the cDNA. The ligated product was then PCR amplified for 10 cycles using the IDT xGen UDI Primers. The resulting libraries were quantified using the Qubit HSDNA assay and TapeStation DNA 1000 assay. Equal molar concentrations were pooled, diluted to 800pM, and sequenced using a 200 cycle NextSeq 2000 flow cell.