GSM8443683: primary tumor; Homo sapiens; RNA-Seq
RNA was isolated using the Qiagen RNeasy kit according to manufacturer's instructions. Using the NEBNext Single Cell/Low Input RNA Library Prep Kit for Illumina (E6420L) and a normalized RNA input between 200pg-50ng a template switching method is used to generate cDNAs. The NEB Ultra II FS workflow is used for fragmentation, end repair, dA-tailing, adapter ligation and PCR. Indexed libraries that meet appropriate cut-offs for both quantity and quality are quantified by qRT-PCR using a commercially available kit (KAPA Biosystems) and insert size distribution determined with the LabChip GX or Agilent Bioanalyzer. Samples with a yield of ≥0.5 ng/ul are used for sequencing.