organism: Mus musculus
description: Protocols: Mouse tissues were collected on PND1 and PND5. Mice were sacrificed with CO2 asphyxiation and organs/tissues were rinsed in saline, snap-frozen in liquid N2 and stored in -80°C until processing. For extraction of RNA samples, tissues were pulverized in liquid N2 with a mortar and pestle and total RNA was purified with RNeasy Mini Kit and treated with RNase-free Dnase I. RNA purity, integrity and concentration were determined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Inc, USA). Only RNA with a RIN value of 1.8 or higher was taken forward for sample preparation and library construction following the manufactory instructions (Illumina Tru Seq RNA sample preparation v2 Guide, Part #15026495 Rev.B February 2012 - “Low sample protocol”). In brief, 1 µg of total RNA from each mouse was taken forward for library preparation. mRNA was purified, chemically fragmented for 8 min at 94oC, and processed for 1st strand synthesis cDNA and then 2nd strand synthesis. After some purification steps, the cDNA was then end-repaired, purified, adenylated at the 3'-ends, and purified before adding the indexed adaptor sequences. TruSeq RNA sample prep kit v2, set A, cat-RS-122-2001 was used for indexing. Each library preparation was then enriched by 10 cycles of PCR, purified and finally validated in regards to size and concentration. For sizing, libraries were analyzed on the Agilent 2100 Bioanalyzer using a DNA 1000 kit from Agilent Technologies.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2016-08-25T17:01:15Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-09T02:29:41Z
attributes.tag: External Id
attributes.value: SAMEA3463460
attributes.tag: INSDC center name
attributes.value: University of Southern Denmark Department of Biochemistry and Molecular Biology
attributes.tag: INSDC first public
attributes.value: 2016-08-25T17:01:15Z
attributes.tag: INSDC last update
attributes.value: 2018-03-09T02:29:41Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-3664:Spinal_cord_PND5_Het_4
attributes.tag: age
attributes.value: 5
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: genotype
attributes.value: heterozygous
attributes.tag: organism part
attributes.value: spinal cord
attributes.tag: sample name
attributes.value: E-MTAB-3664:Spinal_cord_PND5_Het_4
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male