Agilent 2100 bioanalyzer high sensitivity dna analysis kit

RNA yield was determined by Qubit RNA HS assay kit (ThermoFisher Scientific, Reinach, Switzerland). 50 ng of total RNA were subjected to depletion of ribosomal RNA by the use of the RiboMinus™ Eukaryote System v2 kit (ThermoFisher Scientific/Life technologies™/Ambion®, Reinach, Switzerland). As next step, RNA library preparation was achieved by application of the NEXTflex™ Illumina RNA-Seq Library Prep Kit v2 with Molecular Indexes by Bioo Scientific© (Bioo Scientific, Austin, Texas, United States) according to the manufacturer’s recommendations. Library size was assessed before sequencing using the Agilent 2100 Bioanalyzer high sensitivity DNA analysis kit (Agilent, Waldbronn, Germany). Barcoded libraries were sequenced on an Illumina HiSeq 2500 in 75bp paired-end high output mode.

RNA yield was determined using an Agilent 2100 bioanalyzer with the eukaryote total RNA pico assay (Agilent, Waldbronn, Germany). In average 26.8 ± 7.4 ng total RNA were subjected to depletion of ribosomal RNA using the RiboMinus™ Eukaryote System v2 (life technologies, Darmstadt, Germany) according to manufacturers' instructions and directly used for preparation of 10 libraries with the NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® (New England BioLabs Inc., Frankfurt am Main, Germany). Library size was assessed before sequencing using the Agilent 2100 bioanalyzer high sensitivity DNA analysis kit. In total, 6 small RNA libraries were successfully created from in average 84.4 ± 19.1 ng of total RNA with NEBNext® Small RNA Library Prep Set for Illumina® (New England BioLabs Inc., Frankfurt am Main, Germany). Barcoded libraries were sequenced on an Illumina HiSeq 2500.