GSM8118147
5x5mm sections of liver metastasis was isolated to obtain immune cell suspension to perform CD45+ cell sorting. Then, single cells were submitted to squencing. cDNA sequencing libraries were prepared using the Next GEM Single Cell 3' Reagent Kits v3.1 (10X Genomics, PN-1000268) Briefly, after GEM-RT clean up, cDNA was amplified during 12 cycles and cDNA quality control and quantification were performed on an Agilent Bioanalyzer High Sensitivity chip (Agilent Technologies). cDNA libraries were indexed by PCR using the PN-1000215 Dual Index Kit TT Set A. Size distribution and concentration of 3' cDNA libraries were verified on an Agilent Bioanalyzer High Sensitivity chip (Agilent Technologies). Finally, sequencing of cDNA libraries was carried out on an Illumina NovaSeq 6000 using the following sequencing conditions: 28 bp (Read 1) + 10 bp (i7 index) + 10bp (i5 index) + 90 bp (Read 2), to obtain approximately 20-30.000 reads per cell.