title: 2DF-derived iMacs, cell line iMA (rep 2), day 19
design: For RNA isolation, the cells were homogenized using syringe and needle or a QIAshredder kit, and RNA was isolated using RNeasy Micro (<5x105 cells) or RNAeasy Mini (>5x105 cells) Kits. RNA concentrations were measured using a Qubit RNA HS Assay Kit and RNA integrity number (RIN) was assessed using an RNA 6000 Pico Kit for Bioanalyzer. Only RNA samples with RIN > 8 were used for subsequent library preparation. PolyA RNA was isolated using a NEBNext Poly(A) mRNA Magnetic Isolation Module, containing magnetic Oligo d(T) beads. Transcriptome libraries were constructed using a NEBNext Ultra II RNA Library Prep Kit for Illumina following manufacturers instructions. Libraries were prepared with a starting amount of 150 ng and amplified with 15 PCR cycles. RNA was fragmented to the average size of 200nt by incubation at 94 C for 10 min. Libraries were quantified with a Qubit dsDNA HS Kit and the size distribution was determined with a High Sensitivity DNA Kit for Bioanalyzer. Libraries were ~ 350bp in length.
library_name: iMA(rep2)_2DF_d+19