Mus musculus domesticus
Protocols: Sciatic nerves of adult mice were cut or crushed at the sciatic 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.
ENA first public
2017-08-30
ENA last update
2017-03-29
External Id
SAMEA103948149
INSDC center alias
Imperial College London
INSDC center name
Imperial College London
INSDC first public
2017-08-30T17:02:17Z
INSDC last update
2017-03-29T11:14:44Z
INSDC status
public
Submitter Id
E-MTAB-5633:cut7day_RNASeq_3
broker name
ArrayExpress
common name
western European house mouse
developmental stage
adult
organism part
sciatic nerve
sample name
E-MTAB-5633:cut7day_RNASeq_3
strain
C57BL/6J