GSM7911971: exosomes derived from human bone marrow mesenchymal stem cells 3; Homo sapiens; miRNA-Seq
Total RNA was extracted utilizing a RNeasy mini kit (Qiagen, Duesseldorf, Germany). The mRNA concentration and quality were subsequently assessed by a Qubit 3.0 fluorometer (Life Technologies, Gaithersburg, USA) and a Nanodrop One spectrophotometer (Thermo Fisher Scientific Inc., Waltham, USA), respectively. The integrity of the total RNA was evaluated using an Agilent 2100 bioanalyzer (Agilent Technologies Inc., Santa Clara, USA), and only samples with RNA integrity number (RIN) values exceeding 7.0 were selected for subsequent experiments. For the cDNA library construction, approximately 1 μg of total RNA was used, following the manufacturer's instructions for the TruSeq Small RNA sample prep kits (Illumina, San Diego, USA). The procedure encompassed ligation of the RNA to 3ʹ adapters, and the attachment of 5′ adapters to the other end of the RNA molecules. Subsequently, the adapter-ligated RNAs were subjected to RT-PCR and amplified using a low number of cycles. Post-PCR amplification, the libraries were pooled and purified via a PAGE gel.