GSM7594082: 6-Infected Apical-out; Homo sapiens; RNA-Seq
Total RNA was isolated from  iPSC-derived alveolar organoids by using Tri reagent The Illumina Stranded mRNA Prep kit was employed for library preparation, following the manufacturer's reference guide and checklist (Document # 1000000124518 v02). In Summary, 100 ng of total RNA input is used for each library prep. First step, the Oligo(dT) magnetic beads purify and capture the mRNA molecules containing polyA tails. The purified mRNA is fragmented and copied into first strand complimentary DNA (cDNA) using reverse transcriptase and random primers. In a second strand cDNA synthesis step, dUTP replaces dTTP to achieve strand specificity. Then, add adenine (A) and thymine (T) bases to fragment ends and ligate adapters. The resulting products are purified with magnetic beads. Then, add indexes to the anchor-ligated DNA fragments with PCR. The resulting products are purified and selectively amplified for sequencing on an Illumina system.