RNA-Seq of homo sapiens: small cell carcinoma of the ovary-hypercalcemic type (SCCOHT): Sample RNA1
RNA was harvested from four SCCOHT tumors using the Qiagen RNeasy Mini Kit. 10ng of total RNA for each sample was used to generate whole transcriptome libraries for RNA sequencing. Using the Nugen Ovation RNA-Seq System v2, 10ng of total RNA for each sample was used to generate double stranded cDNA, which was subsequently amplified using Nugen's SPIA linear amplification process. Amplified products were cleaned using Qiagen's QIAquick PCR Purification Kit and quantitated using Invitrogen's Quant-iT Picogreen. 1µg of amplified cDNA was fragmented on the Covaris E210 to a target size of 300bp and Illumina's TruSeq DNA Library Preparation Kit with the manufacturer's gel-free protocol was used to generate libraries. Final libraries were evaluated on the Agilent Bioanalyzer, quantitated using the Life Technologies/Invitrogen Qubit, and equimolarly pooled.