Travis K Hughes, Marc H Wadsworth, Todd M Gierahn, Tran Do, David Weiss, Priscila R Andrade, Feiyang Ma, Bruno J de Andrade Silva, Shuai Shao, Lam C Tsoi, Jose Ordovas-Montanes, Johann E Gudjonsson, Robert L Modlin, J Christopher Love, Alex K Shalek
High-throughput single-cell RNA-sequencing (scRNA-seq) methodologies enable characterization of complex biological samples by increasing the number of cells that can be profiled contemporaneously. Nevertheless, these approaches recover less information per cell than low-throughput strategies. To accurately report the expression of key phenotypic features of cells, scRNA-seq platforms are needed that are both high fidelity and high throughput. To address this need, we created Seq-Well S3 ("Second-Strand Synthesis"), a massively parallel scRNA-seq protocol that uses a randomly primed second-strand synthesis to recover complementary DNA (cDNA) molecules that were successfully reverse transcribed but to which a second oligonucleotide handle, necessary for subsequent whole transcriptome amplification, was not appended due to inefficient template switching...
October 13, 2020: Immunity