title: GSM6924804: Hypoxia Monoculture; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was harvested using Rneasy(R) Micro Kit (Qiagen). 20ng/ul of total RNA was used for the construction of sequencing libraries. Sequencing was performed at Single Cell Discoveries, a sequencing service provider. mRNA was processed as described previously, following an adapted version of the single-cell mRNA seq protocol of CEL-Seq [Hashimshony et al. 2012; PMID: 22939981 and Simini et al, 2014; PMID: 25500896]. In brief, samples were barcoded with CEL-seq primers during a reverse transcription and pooled after second strand synthesis. The resulting cDNA was amplified with an overnight In vitro transcription reaction. From this amplified RNA, sequencing libraries were prepared with Illumina Truseq small RNA primers. The DNA library was paired-end sequenced on an Illumina Nextseq™ 500, high output, with a 1x75 bp Illumina kit (R1: 26 cycles, index read: 6 cycles, R2: 60 cycles). located in the Netherlands using an adapted version of the CEL-seq protocol.