GSM5709386: BT474-Tuca-DTP-3_S3; Homo sapiens; RNA-Seq
Samples were prepared by the Princess Margaret Genomic Centre following the 10X Genomics Single Cell 3‚Äô Reagent Kits v2 user guide. Briefly, samples were washed two times in PBS (Life Technologies) + 0.04% BSA (Sigma) and re-suspended in PBS + 0.04% BSA. The cell suspension was loaded onto a 10X single cell A chip. After droplet generation, samples were transferred onto pre-chilled 96-well plates (Eppendorf), heat-sealed, and incubated overnight in a Veriti 96-well thermal cycler (Thermo Fisher) for reverse transcription (RT). Following RT, cDNA was recovered using the Recovery Agent provided by 10X, and subsequently purified by using Silane DynaBead (Thermo Fisher) mix, as outlined by the user guide. Purified cDNA was amplified for 13 cycles before purification on SPRIselect beads (Beckman). Samples were diluted 4:1 (elution buffer (Qiagen):cDNA), and cDNA concentration was determined with a  Bioanalyzer (Agilent Technologies). cDNA libraries were prepared as outlined by the Single Cell 3‚Äô Reagent Kits v2 user guide with modifications to the PCR cycles based on the calculated cDNA concentration.