title: GSM7883161: NHBE_3D_O-3; Homo sapiens; RNA-Seq
library_construction_protocol: Cells/organoids were lysed in RLT buffer, then processed for RNA extraction, or flash frozen, and stored in -80oC until future extraction. RNA was isolated using the Quick-RNA Microprep Kit (Zymo Research) The cDNA was generated using the SMART-Seq® v4 Ultra® Low Input RNA Kit (Takara Bio). The final sequencing library was made using the Nextera XT kit (Illumina). The library construction used Nextera XT chemistry which tagmented 200pg of DNA and added adapters in one step. The fragmented and tagged DNA was then PCR amplified for 12 cycles to incorporate dual-indexing sequencing adapters to generate the final library. A combination of both concentration and library size patterning was used as QC metrics to determine the success of the assay. The UV Qubit fluorometer quantitation step determined the concentration of the library. The Bio/Fragment Analyzer trace revealed the size distribution and concentration of the final sequencing library. Based on these three parameters we assigned library scores to each library. Libraries with a pass score were then used for sequencing in NextSeq 550.