GSM9524092: PBMC Healthy Donor 1 - wild-type E. coli treated; Homo sapiens; RNA-Seq
Total RNA was extracted from cells preserved in DNA/RNA Shield (Zymo Research) according to the manufacturer's instructions. RNA was purified using a bead-based extraction method, quantified using a fluorescence-based assay, and normalized prior to library preparation Libraries were generated using a 3′ mRNA-seq protocol. Poly(A)+ RNA was reverse transcribed using poly(dT)VN primers, followed by second-strand synthesis. Double-stranded cDNA underwent transposase-based tagmentation for adapter incorporation. Libraries were PCR-amplified with UMI-containing unique dual indices and sequenced single-end on an Illumina platform to capture transcript 3′ ends.