title: HiSeq X Ten paired end sequencing; RNA-Seq of four types of mouse immune samples including those of the spleen (SP), bone marrow (BM), lymph node (LN), and peripheral blood mononuclear cell (PBMC)
design: RNA-Seq of four types of mouse immune samples including those of the spleen (SP), bone marrow (BM), lymph node (LN), and peripheral blood mononuclear cell (PBMC)
library_construction_protocol: Six to eight-week-old C57BL/6 mice were purchased from Shanghai SLAC Laboratory Animal Co. All mice were maintained in a barrier facility at Animal Center of SooChow University. To evaluate the immune cell compositions in normal mouse immune organs, 12 mice from four immune organs, including spleen (SP), bone marrow (BM), lymph node (LN) and Peripheral blood mononuclear cell (PBMC), were isolated with routine experimental methods. The isolated sample tissues were soaked in a clean PBS solution, grind into a single cell suspension. Cell suspensions were then filtered by the cell sieves before subjected to a centrifugation at 450 x g for about 10 minutes. Supernatants were discarded and 3 times the suspension volume of Erythrocyte lysate were added into each sample and incubated at room tempreture for 3 minutes. The suspension were then re-suspend with PBS and subjected to a centrifugation at 450xg for about 10 minutes. The cells were finally collected after removing the supernatant. Total RNA was extracted within one week of harvesting cells using the Eukaryotic Poly-A RNA Control Kit  (Affymetrix, USA) according to the manufacturer's instructions. A NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB) were used for rRNA depletion. Complementary DNA (cDNA) library preparations and sequencing were carried out in Suzhou Institute of Systems Medicine. The E6310L NEBNext Ultra RNA Library Prep Kit for Illumina（NEB, E7530S）(NEB) was used according to the manufacturer's instructions.
attributes.tag: Experimental Factor: cell type
attributes.value: bone marrow cell