Total RNA sequencing for Mus musculus bone marrow lymphoid committed cell populations

MPP2, MPP3 and CLP populations were isolated by fluorescence-activated cell sorting (FACS) from the BM of B6 mice based on the surface expression of VCAM1, FLT3 and IL7Ra within LSK population. On average 100K cells were analyzed for each population in 3 independent biological experiments. We generated sequence libraries from these low RNA inputs using the SMARTer Stranded RNA-Seq and sequenced at high depth of coverage to generate ~2x120 M reads per biological replicate.MPP2、MPP3及CLP细胞群通过荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）从B6小鼠的骨髓（Bone Marrow, BM）中分离得到，分选标准为LSK细胞群内血管细胞黏附分子1（VCAM1）、FMS样酪氨酸激酶3（FLT3）与白细胞介素7受体α链（IL7Ra）的表面表达水平。每个细胞群平均采用10万个细胞进行分析，共完成3次独立生物学重复实验。本研究针对上述低起始量RNA样本构建测序文库，所用技术为SMARTer Stranded RNA-Seq，并以高测序覆盖度开展测序，每个生物学重复可获得约2×120 M条测序读段（reads）。