GSM8753119: AZD5991_Mouse0_Tumor_Plate1; Homo sapiens; RNA-Seq
Single cells were sorted into a 96-well plate with each well containing 1% 2-mercaptoethanol and 10 µL of TCL buffer (Qiagen). RNA purification was performed using RNA-SPRI beads (Beckman) prior to cDNA synthesis. Full-length single cell RNA-Seq libraries were prepared using the Smart-Seq2 protocol. Single cells were sorted into a 96-well plate with each well containing 1% 2-mercaptoethanol and 10 µL of TCL buffer (Qiagen). RNA purification was performed using RNA-SPRI beads (Beckman) prior to cDNA synthesis. RNA was reverse-transcribed using Maxima RNAse H-minus (ThermoFisher) in the presence of biotinylated oligo-dT30VN (/5BiosG/ AAGCAGTGGTATCAACGCAGAGTACT(30)VN), biotinylated template-switching oligonucleotides (/5BiosG/AAGCAGTGGTATCAACGCAGAGTACATrGrG+G), and betaine. cDNA amplification was performed using 2x KAPA HiFi Hotstart Ready Mix (Roche) and biotinylated ISPCR primers (/5BiosG/ AAGCAGTGGTATCAACGCAGAGT). Tagmentation was performed on 0.15 ng of amplified cDNA using the Nextera XT kit (Illumina) and amplified with Nextera indexes. Smart-Seq2 libraries were sequenced paired-end on an Illumina NextSeq500 sequencer.