GSM9206948: Immune cells, H_1; Mus musculus; RNA-Seq
Tumors were subjected to single cell isolation. Tumor-infiltrating immune cells were isolated by using Percoll and gradient centrifugation. Single cells were isolated into oil emulsion droplets with barcoded gel beads and reverse transcriptase mix using the DNBelab C-TaiM 4 (MGI), cDNA was generated within these droplets, then the droplets were dissociated. cDNA was purified. cDNA amplification was performed by PCR using reagents within the DNBelab C High-throughput single-cell RNA library preparation kit V3.0（MGI） Amplified cDNA was purified using SPRIselect magnetic beads (Beckman Coulter) according to the protocol. cDNA was fragmented, end repair and A tailing, the DNAs were ligated to dual Adaptor (DNBelab C Single Cell Library Preparation Sample Labelling Kit S). The ligated products were amplified and purified, quantified (Qubit 4.0) and stored at -80℃ before sequencing. For high-throughput sequencing, the libraries were prepared following the manufacturer's instructions and applied to DNBSEQ-T7 system for 150 nt/100 nt paired-end sequencing.