GSM3664952: Undiff_b; Homo sapiens; RNA-Seq
Total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies, Inc.) The RNA was quantified (Qubit RNA Assay Kit, Life Technologies, Inc.), and quality controlled (RNA6000 Nano Kit and BioAnalyzer 2100, Agilent). 500 ng of total RNA was used as input for the Illumina TruSeq Stranded mRNA Sample Prep Kit (Illumina, Inc.) and sequencing libraries were generated according to the manufacturer's protocol. Briefly, the ribosomal RNA was depleted using RiboZero Gold (Illumina, Inc.), followed by a fragmentation step. The fragmented RNA was then copied into first strand cDNA using random primers and reverse transcriptase. Next, second strand cDNA synthesis was achieved using DNA Polymerase I and RNase H. The cDNA was then ligated to Illumina supplied adapters and enriched with PCR to create the final cDNA libraries.