characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-08-30T17:02:17Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-29T11:14:44Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103948151
characteristics.characteristic: External Id
characteristics.text: Imperial College London
characteristics.characteristic: INSDC center name
characteristics.text: 2017-08-30T17:02:17Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-29T11:14:44Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1637319
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5633:uncut_RNASeq_2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: western European house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: No injury was performed on control samples. Sciatic nerves of adult mice were harvested at the sinotic notch. RNA was extracted from samples using Trizol (Invitrogen) according to manufacturer’s instructions with an additional purification step by on-column DNase treatment using the RNase-free DNase Kit (Qiagen) to ensure elimination of any genomic DNA. The integrity and quantity of RNA was determined using Qubit (Invitrogen) and Agilent 2100 Bioanalyzer (Agilent Technologies). 1 µg of total RNA was used to generate RNA-Seq libraries using TruSeq Stranded total RNA sample preparation kit (Illumina) according to the manufacturer’s instructions. Briefly, cytoplasmic and mitochondrial rRNA was removed using biotinylated, target specific oligos combined with Ribo-Zero rRNA removal beads. Following purification, RNA was fragmented using divalent cations under elevated temperature and cleaved fragments were copied onto first strand cDNA using reverse transcriptase and random primers. Next, the second strand was synthesised using DNA Polymersae I and RNase H. cDNA 3’ ends were adenylated and indexed adapters ligated followed by 10 cycles of library amplification. Finally, the libraries were purified using AMPue XP Beads (Beckman Coulter) and quality was evaluated using Agilent 2100 Bioanalyzer. Three libraries were run on a single lane of the HiSeq 2000 platform (Illumina) to generate 100 bp paired-end reads.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Mus musculus domesticus
characteristics.characteristic: organism
characteristics.text: Mus musculus domesticus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10092
characteristics.characteristic: organism
characteristics.text: sciatic nerve
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0001322
characteristics.characteristic: organism part
characteristics.text: Mus musculus domesticus
characteristics.characteristic: scientific_name
characteristics.text: C57BL/6J
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0000606
characteristics.characteristic: strain
characteristics.text: uncut_RNASeq_2
characteristics.characteristic: title