GSM2339475: Hypothalamus, BXD43; Mus musculus; RNA-Seq
Total RNA extraction from hypothalamus and brainstem samples was performed using the peqGOLD TriFast method (Peqlab, Erlangen, Germany) (Mounien et al., 2010). RNA integrity was verified with a 2100 Bioanalyser (Agilent, Santa Clara, USA). For each BXD strain, equal amounts of RNAs from 3 to 6 hypothalami were pooled in a final volume of 50 μL RNA-seq libraries were prepared using 1µg of pooled RNAs and the Illumina TruSeq Stranded mRNA reagents (Illumina; San Diego, California, USA) on a Sciclone liquid handling robot (PerkinElmer; Waltham, Massachusetts, USA) using a PerkinElmer-developed automated script. Cluster generation was performed with the resulting libraries using the Illumina TruSeq SR Cluster Kit v3 reagents and sequenced on the Illumina HiSeq 2000 using TruSeq SBS Kit v3 reagents. Sequencing data were processed using the Illumina Pipeline Software version 1.82. A sequencing depth of 40 millions of 100-bp single-end reads was used.
GEO Accession
GSM2339475