title: GSM7211228: ATlo oHSC_3; Mus musculus; RNA-Seq
library_construction_protocol: RNA was isolated from ~10,000 HSCs using the RNeasy Plus Micro Kit (Qiagen, 74034). RNA integrity number (RIN) was determined by Bioanalyzer (Agilent Technologies) and RNA samples with RIN > 9.0 were further processed. RNA samples were processed over two experimental “sort days” and stored at -80°C in RNA isolation buffer until library preparation. A total of 3 paired AThi/ATlo oHSC biological replicates were sequenced alongside 2 yHSC replicates. RNA was quantified and QC using an Agilent Bioanalyzer. Illumina sequencing libraries were prepared and sequenced at the NYU Genome Technology Center (New York, NY). The NuGen Ovation Trio low input RNA-seq library preparation kit was used for library preparation. All libraries were prepared at the same time in order to minimize batch effects. Libraries were sequenced using an Illumina HiSEQ 4000 instrument with paired end 50 base pair sequencing to a target depth of 40 million unique reads per sample.