GSM9540422: CD45+ cell, BiDT; Mus musculus; RNA-Seq
Tumor tissues were collected, cut into small pieces, and digested with 100μg/ml DNase I (Roche) and 1 mg/ml collagenase IV (Roche) at 37 °C for 40 min. The digested tumors were then passed through a 70 μm cell strainer to make single-cell suspensions. Single cells were blocked with anti-CD16/32 Ab (clone 93) for 30 min and then stained with specific antibodies for 30 min on ice. Live CD45+ cells were sorted by FACS, and five biological replicates were amalgamated for library construction. The sorted cells were loaded onto a Chromium single-cell chip to generate barcoded nucleic acid fragments. Single-cell transcriptomic libraries were generated using the commercial workflow specified by 10x Genomics (https://cdn.10xgenomics.com/image/upload/v1660261285/support-documents/CG000204_ChromiumNextGEMSingleCell3_v3.1_Rev_D.pdf).