GSM6256759: 643; Mus musculus; RNA-Seq
otal RNA was extracted from total BM at the indicated time points using RNeasy micro kit (Qiagen) according to the manufacturer's instruction. The quality of the isolated total RNA from each sample was checked on 2200 Tapestation System (Agilent, Santa Clara, USA) using the RNA ScreenTape assay and the quantitation was performed using the NanoVue Plus Spectrophotometer (Biochrom Ltd, Cambridge, UK). 100-300 ng of total RNA per sample with a RNA integrity number >8 were used for library preparation. The RNA-Seq library preparation was performed using TruSeq Stranded mRNA Sample Preparation kit (Illumina, San Diego, Calif) according to the manufacturer's instructions (High Sample Protocol). The RNA-Seq libraries were checked on the 2200 Tapestation System (Agilent) using the High-Sensitivity DNA 1000 ScreenTape.assay. Libraries were quantified by qPCR on a LightCycler 480 System (Roche Molecular Diagnostics, Pleasanton, Calif) using KAPA Library Quantification Kit for Illumina platforms (Kapa BiosysTem cells, Boston, Massachusetts). Libraries were then pooled in equal quantities, checked and quantified using same techniques (2200 Tapestation and LightCycler 480). Paired-end sequencing (2 x 150 bps) of these libraries was performed on a  Novaseq 6000 S4 Illumina sequencing using NextSeq 500 High Output kit v2 (Illumina) generating in average 20 million pairs of reads per sample.