title: Illumina HiSeq 2500 sequencing; Expression profiling of adult mouse brain and liver tissues (RNA-seq)
design: Expression profiling of adult mouse brain and liver tissues (RNA-seq)
library_construction_protocol: Wild-type male and female mice at 3 months of age were purchased from National Laboratory Animal Center and maintained in pathogen-free facility. Mice were sacrificed with five brain sub-regions (cortex, hippocampus, thalamus, hypothalamus, and cerebellum) and liver being harvested. The animal protocol was approved by the Institutional Animal Care and Use Committee of the Taipei Medical University. Tissues were homogenized in the TRIzol reagent (Thermo Fisher Scientific). Total RNA was extracted twice with phenol-chloroform and precipitated with isopropanol. Subsequently, in order to remove the chemicals, total RNA was precipitated with ethanol twice and dissolved in autoclaved water. 1 microgram of total RNA with RIN (RNA integrity number) ranged from 8.0 to 10 was subjected to the removal of rRNA using Ribo-Zero Gold kit for human/mouse/rat (Epicentre, Madison, WI). The sequencing library for rRNA-depleted RNA was prepared using TruSeq Stranded mRNA Sample Prep Kit (Illumina) which produced directional RNA-seq libraries. The adapter-ligated double-stranded cDNA library was enriched with 6 to 7 cycles of PCR using KAPA HiFi DNA polymerase.
attributes.tag: Experimental Factor: thalamus
attributes.value: organism part
attributes.tag: Experimental Factor: female
attributes.value: sex