title: GSM2805860: exHSC_rep1; Mus musculus; RNA-Seq
library_construction_protocol: Cells were sorted into Trizol-LS. Total RNA was prepared using RNeasy micro kit (Qiagen, Valencia, CA, USA). DNase treatment was performed either on-column (Qiagen) or using DNA-free DNA Removal Kit (Life Technologies). Total RNA was quantified with RNA HS Kit (Q32852) for Qubit fluorometer (Life Technologies) and analyzed for integrity using RNA 6000 Pico Kit (5067-1513) for 2100 Bioanalyzer (Agilent). mRNA was isolated from total RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490, NEB). PolyA-selected mRNA was fragmented to average size of 300 nt, reverse transcribed to generate double-stranded cDNA, and converted to a paired end library using NEBNext Ultra RNA Library Prep Kit for Illumina (E7530, NEB) according to manufacturer's instructions including the optional double size selection procedure using Agencourt AMPure XP beads. Prepared libraries were quantified with dsDNA HS Kit (Q32851) for Qubit and the size distribution was assessed using High Sensitivity DNA Kit (5067-462) for Bioanalyzer. Libraries were sequenced on Illumina HiSeq2500 in paired-end mode with the read length of 75 nt.
attributes.tag: GEO Accession
attributes.value: GSM2805860