GSM8744806: A12; Homo sapiens; RNA-Seq
Fresh tumor samples were transported expeditiously to the Singleron lab. The tumor tissues were washed three times with Hanks Balanced Salt Solution (HBSS), cut into 1e2 mm pieces, and digested in 2 ml GEXSCOPE® Tissue Dissociation Solution (Singleron) at 37 。C for 15 min with constant but gentle agitation. The digested samples were ﬁltered using 40-micron  sterile  strainers  and  centrifuged  at  800×g for 5   min.  The   pellets  were   then   resuspended   in   1   ml phosphate-buffered saline  (PBS;  HyClone, United States), and 2 ml GEXSCOPE® red blood cell lysis buffer (Singleron) was added at 25 。C for  10 min to  remove the  red blood cells. The tumor cell-containing solution was centrifuged at 500×g for 5 min, and the cells were resuspended in PBS. The samples were stained with trypan blue (Sigma, United States) to verify cell viability. Single-cell suspensions with 1  × 105  cells/mL in  PBS were prepared. Then, the suspensions were loaded onto micro- ﬂuidic devices, and scRNA-seq libraries were constructed according  to  the  Singleron  GEXSCOPEⅢ  protocol  in  the GEXSCOPEⅢ  Single-Cell  RNA  Library  Kit  (Singleron  Bio- technologies).21   Individual  libraries were diluted to 4  nM and  pooled for sequencing.  Pools were sequenced on an Illumina HiSeq X with 150 bp paired-end reads.