GSM8515669: MSC 351 3D; Homo sapiens; RNA-Seq
whole RNA was isolated by the Nucleo Spin RNA Kit (Macherey & Nagel) for samples from Donor 125 and 135: The indexed cDNA library was generated using the TruSeq Stranded mRNA LT Sample Prep Kit 24 (Illumina), according to the TruSeq Stranded mRNA Sample PreparaCon Protocol LS (Illumina). The inital quantity of total RNA was 800 ng for bone marrow stromal cells for samples from donor 351 and 355: Total RNA samples were quantified using a Qubit Fluorometer, and RNA integrity was checked on a TapeStation (Agilent). Double-indexed stranded mRNA-Seq libraries were prepared using the ILMN Stranded mRNA Library Prep Kit (Illumina, #20040534), starting from 500 ng of input material according to the manufacturer's instructions. for samples from donor 125 and 135: Libraries were quality checked by qPCR and equimolarly pooled based concentration measurements (nanodrop). The sequencing was performed on the Illumina MiSeq System. for sampls from donor 351 and 355: Libraries were equimolarly pooled based on Qubit concentration measurements and TapeStation size distributions. The loading concentration of the pool was determined using a qPCR assay (Roche, #7960573001). Libraries were then sequenced on the Illumina NovaSeq X Plus platform using PE100 sequencing mode, with a target of 50 million reads per library.