GSM7082513: cells_chip1_mRNA; Homo sapiens; RNA-Seq
Cultured cancer cells were harvested and dissociated, fresh cells were resuspended with 1 x Dulbecco's phosphate-buffered saline (DPBS, C14190500BT, Thermo Fisher Scientific).  Mice were euthanized using carbon dioxide and lung tissues were collected and cut into small pieces of 2-4 mm. These pieces were dissociated into single-cell suspensions with PythoN® Tissue Dissociation System following the manufacturer's instructions (MD1101001, Singleron Biotechnologies). Single-cell suspensions were dispensed into the ICELL8 nanowell chip for cell imaging and screening. Selected single live cells were lysed to release total RNA; then, small RNAs were ligated with 3' adapters, and mRNAs were reverse transcribed and linked to adapters using template switching oligos. Next, the remaining 3' adapters were removed, and 5' adapters were ligated to small RNAs. The ligated small RNAs were then reverse transcribed. Cell barcodes were introduced by PCR to label individual cells (PCR-1). Next, small RNA and mRNA libraries were separately collected according to their size ranges. Then, the libraries were amplified for high-throughput sequencing (PCR-2). Additionally, by-products in small RNA libraries were removed. PSCSR-seq V2