GSM8741817: Teratoma_CD45_Rep2; Homo sapiens; RNA-Seq
For 10× genomics, cells were counted and then loaded into Chromium microfluidic chips with 30 (v2 or v3, depends on project) chemistry and barcoded with a 10× Chromium Controller (10X Genomics). RNA from the barcoded cells was subsequently reverse-transcribed. For MobiNova-100, CD8+ T cells were collected and extracted following the manufacturer's instructions (MobiDrop, Shanghai Baimu Biotechnology Co., Ltd.). After quality control assessment, samples were loaded to a pre-prepared chip containing reverse transcription reagents, cell-phase reagents, and droplet generation oil. The chip holder was then carefully transferred to the MobiNova®-100 instrument for droplet generation and single-cell barcoding. A droplet pre-processor was employed to pre-treat the generated droplets, before reverse-transcription to cDNA. The resultant cDNA after reverse transcription underwent GEM breaking and pre-amplification. Subsequently, SPRIselect beads were used for cDNA purification, and the cDNA quality was assessed using Qubit and Agilent 2100 HS DNA. For 10× genomics, sequencing libraries were constructed with reagents from a Chromium Single Cell 30 v2(v2 or v3, depends on project) reagent kit (10X Genomics) according to the manufacturer's instructions. Sequencing was performed with Novaseq xplus according to the manufacturer's instructions (Illumina). For MobiNova-100, library construction included the following steps: cDNA fragmentation, adapter ligation to the fragmented products, size selection of the ligated products, amplification of the library with tags, and size selection of the amplified library fragments.