title: GSM4781672: NSG_CT_1; Mus musculus; RNA-Seq
library_construction_protocol: RNA extraction was performed using standard TRIzol / chloroform extraction followed by 100% ethanol precipitation and RNA purification using the RNeasy Mini Kit with DNase treatment. Total RNA was quantified using a Nanodrop 1000 spectrophotometer and qualified with an Agilent 2100 Bioanalyzer using the Agilent RNA 6000 Nano Kit according to the manufacturer's instructions. Libraries from total RNA were prepared using the TruSeq®Stranded Total RNA Gold Library Preparation Kit according to the manufacturer's protocol. Briefly, 0.5 µg of total RNA was ribo-depleted using the Ribo-Zero Gold Kit. RNA fragmentation resulted in fragments of 80 – 450 nt, with a major peak at 160 nt. First-strand cDNA synthesis by random hexamers and reverse transcriptase was followed by second-strand cDNA synthesis, performed in presence of dUTP instead of dTTP. Blunt-ended double-stranded cDNA was 3´adenylated and Illumina-indexed adapters were ligated. Resulting libraries were enriched with 15 PCR cycles.