GSM7915077: bone marrow mix of wildtype and Ythdf2 cKO mice; Mus musculus; RNA-Seq
BM cells were collected and treated one time with 10 mL ammonium chloride (07850, StemCell technologies) followed by two times wash with 2% regular RPMI 1640 cell culture medium. Then, diluted 1 million cells of each BM sample in 49.5 µL of cell staining buffer (420201, Biolegend) in 12 x 75mm flow cytometry tubes and blocked cells with 0.5 µL anti-mouse CD16/CD32 antibody for 10 minutes on ice. Then diluted 1 µg TotalSeq type C antibody into cell staining buffer with a final volume of 50 µL and centrifuged at 14, 000 g at 4ºC for 10 minutes and subsequently added into 50 µL blocked cell suspension and incubated 30 minutes at 4ºC. Each antibody stained one mouse BM cells. After staining, a total wash of 3 times with 3 mL cell staining buffer by spinning down at 4ºC for 5 minutes at 600 g followed by filtering cells through a 40 µm Flowmi Cell strainer (BAH136800040-50EA, Sigma-Aldrich) with 500 mL cell staining buffer. Combined the 4 mice BM cells with featured antibodies with equal cell amounts. Then prepared a total number of exactly 10, 000 cells for 10×Genomics single cell platform to generate single cell by partitioning and recovery within the 10× Chromium chip and prepare the library. Chromium Next GEM Single Cell 5' Reagent Kits v2 with Feature Barcode technology for cell surface protein