GSM8678735: Aorta_pericyte_monolayer_Day1; Homo sapiens; RNA-Seq
Monolayer cells and spheroids formed on nanofiber scaffolds were collected in Buffer RA1 (Takara Bio) +1% beta-mercaptoethanol (Fisher Scientific). NucleoSpin RNA kit from Takara Clontech was used for RNA extraction, 50ng input RNA used for library construction. RNA was assessed for quality using an Agilent TapeStation 4150/Fragment Analyzer 5300 and RNA concentration was quantified on a Qubit FLEX fluorometer. Libraries were generated with the Illumina Stranded Total Library Prep kit (Illumina: 20040529) according to the manufacturer's instructions. Briefly, 6 ng of input RNA was used for each sample. Following adapter ligation, 15 cycles of indexing PCR were completed, using Illumina RNA UD Indexes. Library quantification and assessment was done using a Qubit FLEX fluorometer and an Agilent TapeStation 4150/Fragment Analyzer 5300. Libraries were normalized and pooled to 2 nM by calculating the concentration based off the fragment size (base pairs) and the concentration (ng/ml) of the libraries. Sequencing was performed on an Illumina NextSeq 2000, using a P3 flow cell. The pooled library was loaded at 750 pM and sequencing was carried out with read lengths of 2x101 bp, with a target of 40 million reads per sample. Sequencing data was demultiplexed by the on-board Illumina DRAGEN FASTQ Generation software.