title: RNA sequencing: Sample LCLF-005_02
design: Stranded cDNA sequencing libraries were generated with a TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA) following the manufacturer's instructions. Briefly, 500 ng of total RNA was used for mRNA selection. After the reverse transcription to first strand cDNA, strand information was maintained with dUTP during second strand synthesis. A single nucleotide (containing adenine) was added to the dsDNA fragments and the products were ligated to an adapter. The products were then purified and amplified by PCR to create the final cDNA library. The library was qualified with an Agilent Bioanalyzer and quantified with a Qubit 2.0 fluorometer. The cluster generation and pair-end (2 × 75 bp) sequencing were performed on an Illumina HiSeq 3000. Up to 96 barcoded samples were pooled for one single run, which yielded at least 30 M passed filter paired reads per sample. Sequencing results were demultiplexed and converted to FASTQ format using Illumina bcl2fastq software.