GSM4477966: Calu3_totalRNA-untr-4h-A; Homo sapiens; RNA-Seq
For polyA/total/small RNA-seq, cells were lysed in Trizol and RNA extracted using standard methods. For single-cell RNA-sequencing, cells were fixed in methanol and stored at -80 degrees until further processing. Rehydration was done as described in Alles et al. 2017 (doi: 10.1186/s12915-017-0383-5) PolyA and total RNA-seq libraries were constructed using the NEB Ultra II Directional RNA Library Prep Kit. SmallRNA-seq libraries were constructed using the Clontech SMARTer smRNA-Seq Kit. Single-cell libraries were constructed using a a Nadia device (Dolomite Bio) according to version 1.8 of the manufacturer's protocol, except that the second-strand synthesis step as described in Hughes et al. 2019 (doi: doi.org/10.1101/689273) was included after the exonuclease step.