title: GSM7063590: SS007; Mus musculus; RNA-Seq
library_construction_protocol: Animals were euthanized by IP injection of Euthasol (diluted with  sterile saline solution) Blood samples were  collected immediately after euthanasia by cardiac puncture in a sterile  hood. Blood (400–500 ll) was added to PAXgenet RNA solution  (Becton, Dickinson and Co., Franklin Lakes, NJ) at a 1:5 ratio and  inverted _x0003_10 times to mix. Samples were stored at –80C for the  length of the study. RNA samples were prepared  using the PAXgene protocol, globin-depleted using the mouse  Ambiont GLOBINclear kit (Ambiont; Thermo Fisher Scientifice  Inc., Waltham, MA), and quantified using a NanoDrope One  spectrophotometer (Thermo Fisher Scientific). RNA quality was  determined using the Agilent 2100 Bioanalyzer (Santa Clara, CA), and  RIN values ranged from 8.0 to 9.5 across all samples. RNA sequencing analysis for gene expression was performed according to standard RNA-seq pipeline at the Columbia University Medical Center Genome Center. We used the RIBOZERO workflow in which ribosomal depletion was performed to remove rRNAs from globin cleared total RNA. RINs of globin cleared RNA (catalog number AM1981 from Thermofisher) were in the acceptable range for 40-million reads RNAseq (75% of samples had RIN values >7.5). After rRNA depletion, library construction was performed using Illumina TruSeq Stranded Total RNA kit.