Ovation solo rna seq system human kit

Total RNA was extracted from FACS-sorted cells using an RNeasy mini Kit (Qiagen, Hilden, Germany). The sequencing library was constructed by following the Ovation SoLo RNA-seq system Human kit (NuGEN Technologies, San Carlos, CA, USA), using 5 ng of total RNA to generate cDNA. The cDNA libraries were sequenced by 50-base single-read sequencing on an Illumina NovaSeq 6000 system (Illumina, San Diego, CA, USA). The sequencing run and base call analysis were performed according to the Ovation SoLo RNA-Seq system Human M01406 v4. Raw sequence data were generated with processing by CASAVA-1.8.4 with version RTA 1.17.20.0.

Total RNA was isolated and purified from sorted cells using an RNeasy Micro Kit (Qiagen, Hilden, Germany). RNA-seq libraries were generated with the Ovation SoLo RNA-Seq System, Human kit (NuGEN, Redwood City, CA, USA) using 5 ng of total RNA. The cDNA libraries were sequenced by 50-base single-read sequencing on an Illumina HiSeq 2500 sequencer (Illumina, San Diego, CA, USA). The sequencing run and the base call analysis were performed according to the HiSeq 2500 System Guide with TruSeq SBS kit v3-HS. Raw sequence data were generated with processing by CASAVA-1.8.4 with version RTA 1.17.20.0. Reads were mapped to the hg38 genome with Tophat2. Normalized FPKM values and differential gene expression analyses were generated with Cuffdiff2. Q values (the FDR-adjusted p value after Benjamini–Hochberg correction for multi-testing) lower than 0.05 were considered significant.