scRNA-Seq of mus musculus: tumor
Following cold PBS perfusion, mouse tumor tissue was minced and enzymatically dissociated at 37C for one hour. The resulting cell suspension was filtered and centrifuged. The pellet was then subjected to density gradient centrifugation using a Percoll solution. After centrifugation, the supernatant was discarded, and the cell pellet was treated with a lysing solution to remove erythrocytes. The cells were washed, resuspended in PBS, and counted. Only samples with cell viability exceeding 90% were processed further. Single-cell suspensions were loaded onto a microfluidic chip, and libraries were constructed following the manufacturer's protocol for subsequent sequencing on an Illumina HiSeq 4000 platform.