GSM6890031: hiPSC-CM TKI cardiotoxicity; Homo sapiens; RNA-Seq
Total RNA was isolated using MagMAX™-96 total RNA isolation kit. RNA was transferred into 384-well plates and the subsequent library preparation was done according to this publication(https://pubmed.ncbi.nlm.nih.gov/30333485/) and using an automated liquid dispensing system. RNA was reverse transcribed using the Maxima H Minus Reverse Transcriptase (Thermo Fisher) and barcoded primers containing poly(T) sequence about 24 bases long, a random 10-nucleotide sequence as unique molecular identifiers, a random 6-nucleotide sequence as well barcodes and a sequence complimentary to pre-amplification primers. The other end of cDNA sequences were filled with a sequence complementary to pre-amplication primers through a template switching oligo and the same reverse transcriptase. The cDNAs of different drug treatments were pooled and amplified by PCR. In this experiment, the 10-nucleotide unique molecular identifiers were used to label different RNA species and the 6-nucleotide well barcodes were used to label the 129 TKI treatments, not single cells.