GSM3534976: Ctrl_ICGScluster2_042_Hematopoietic stem cells (CD31+kit+CD45-); Mus musculus; RNA-Seq
CD31+kit+CD45- cells were FACS-purified. Cells were sorted in 2.3 µl of lysis buffer containing 0.2% Triton X-100 (Sigma-Aldrich) and 1U of Superase-In RNase Inhibitor (Ambion) and processed following the Smart-Seq2 protocol. Following preamplification, all samples were purified using Ampure XP beads (Beckman Coulter) at a ratio of 1:0.6. The cDNA was then quantified using the Quant-iT PicoGreen dsDNA Assay Kit (Thermo Fisher) and size distributions were checked on high-sensitivity DNA chips (Agilent Bioanalyzer). Libraries were prepared using the Illumina Nextera XT DNA preparation kit. Single-cell RNA-seq analysis was performed as described previously (Picelli et al., 2014). Pooled libraries were run on the Illumina Hi-Seq 4000. Samples were used to construct Nextera XT libraries (Illumina) from 125 pg of preamplified cDNA. Libraries were purified and size selected (0.5X-0.7X) using Ampure XP beads. Then, libraries were quantified using KAPA qPCR quantification kit (KAPA Biosystems).
GEO Accession
GSM3534976