GSM4985091: single-cell RNAseq of SW480 tumors at 1dpi (library 8); Homo sapiens; RNA-Seq
Single-cell mRNA sequencing was performed at Single Cell Discoveries (single-cell sequencing provider, the Netherlands) according to an adapted version of the SORT-seq protocol (Muraro et al. 2016 with primers described in van den Brink et al. 2017). In short, single cells were FACS sorted into 384-well plates containing 384 primers and Mineral oil (Sigma). After sorting, plates were snap-frozen on dry ice and stored at -80° C. For amplification cells were heat-lysed at 65° C followed by cDNA synthesis using the CEL-Seq2 protocol (Hashimshony et al. 2016) and robotic liquid handling platforms. After second strand cDNA synthesis, the barcoded material was pooled into libraries of 384 cells and amplified using IVT. Following amplification, the rest of the CEL-seq2 protocol was followed for preparation of the amplified cDNA library, using TruSeq small RNA primers (Illumina). 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)