title: GSM4459618: g1.S17; Homo sapiens; RNA-Seq
library_construction_protocol: Individual cell lines, cell line pools prior to injection, and cells isolated from metastases were subjected to RNA-Seq. RNA extraction was performed using Quick-RNA MicroPrep according to instructions (Zymo Research). RNA was quantified using RNA 6000 Pico Kit on a 2100 Bioanalyzer (Agilent). RNA samples from cell numbers lower than 500 were not measured but all were used as input for library preparation. cDNA was synthesized using Clontech SmartSeq v4 reagents from up to 2 ng RNA input. Full length cDNA was fragmented to a mean size of 150bp with a Covaris M220 ultrasonicator and Illumina libraries were prepared from 2 ng of sheared cDNA using Rubicon Genomics Thruplex DNAseq reagents. The finished dsDNA libraries were quantified by Qubit fluorometer, Agilent TapeStation 2200, and RT-qPCR using the Kapa Biosystems library quantification kit. Uniquely indexed libraries were pooled in equimolar ratios and sequenced on Illumina NextSeq500 runs with paired-end 75bp reads.