title: GSM8527251: M1001,scRNA-seq; Homo sapiens; RNA-Seq
library_construction_protocol: Ensure that the volume of colorectal cancer tissue samples is adequate, fresh tissue samples should be ≥200mg, and use pre-cooled PBS or saline to wash tissue samples to remove blood stains and impurities. Remove fat, connective tissue or necrotic tissue around the sample and cut the tissue into small pieces of about 2mm³. Submerge the processed tissue samples in pre-cooled tissue preservation solution and maintain a 4°C environment. The samples were transported to the laboratory for single-cell sequencing experiments within 48 hours. In the laboratory, the tissue samples were cut up and preserved using tissue preservation solution, followed by single cell suspension preparation. Library was performed accroding to manufacturer's instructions (Illumina NextSeq 500,10X Genomics).Briefly, high quality cells were isolated from colorectal cancer tissues to ensure cell viability and purity. Individual cells are lysed and total RNA is extracted.RNA is converted to cDNA using reverse transcriptase, often with primers bearing cell-specific barcodes and unique molecular identifiers (UMIs). Amplify the cDNA by PCR or other methods to increase coverage during sequencing. The amplified cDNA is fragmented and sequencing junctions are added to construct libraries that can be used for high-throughput sequencing. Use a bioanalyser to detect the size distribution and concentration of the library. The library was loaded onto the sequencing platform for high-throughput sequencing.