GSM3732530: L66A; Bos taurus; RNA-Seq
Total RNA was extracted using the PAXgene® Blood miRNA Kit (PreAnalytiX GmbH) according to the manufacturer's protocol. Total RNA was eluted in a final volume of 80 μL. RNA concentration was measured by NanoDrop™ 1000 spectrophotometer (Thermo Scientific) and RNA integrity was assessed with an Agilent 2200 TapeStation system (Agilent Technologies). Libraries were prepared with the Illumina Truseq RNA sample prep kit (Illumina Inc., USA) following the manufacture's protocol and the size and yield evaluated using an Agilent TapeStation 2200. Libraries were then quantified with an ABI9700 qPCR instrument using the KAPA Library Quantification Kit in triplicate, according to the manufacture's protocol (Kapa Biosystems, Woburn, MA, USA) and then normalized to 10 nM as recommended by Illumina for cluster generation on the Hiseq2000. Fifteen libraries were prepared and equimolar amounts of 5 samples were mixed before NaOH denaturation. Each of the pools was run in a lane of a Hiseq Flowcell.
GEO Accession
GSM3732530