title: GSM1651911: WT2_mRNA; Mus musculus; RNA-Seq
library_construction_protocol: 24,000 Flt3- LSK cells were sorted into Trizol and RNA was purified (Qiagen). Spike-in RNA (ERCC RNA Spike-In Mix, Life Technologies) was added following the manufacturer’s instructions. Two types of sequencing libraries were prepared; one from polyA-selected RNA to analyze mRNAs, and one from total RNA to compare rRNA levels. 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