GSM8508882: WT, replicate 1, scRNAseq; Mus musculus; RNA-Seq
Freshly collected femur and tibia from Tbc1d23ΔE15/ΔE15 or wild type mice were cuted in the middle and instantaneous centrifugated to obtained the whole bone marrow cells. The Lin- cells were isolated from whole bone marrow cells by  EasySep™ Mouse Hematopoietic Cell Isolation Kit (STEMCELL Technoligies). Lin- cells were washed and resuspended in PBS buffer without calcium and magnesium. Library was performed according to the manufacter's instructions (Chromium Next GEM Single Cell 3' Solution v2 protocol,10x Genomics). Brifly，bone marrow Lin- cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM).The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an illumina R1 primer sequence. Unique Molecular identifiern (UMI) and 10x barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRlselect reagent for subsequent library construction. During the library construction illumuna R2 primer sequence, paried-end constructs with P5 and P7 sequences and a sample index were added.