characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-04-01T00:23:18Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-04-01T00:23:18Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA111519294
characteristics.characteristic: External Id
characteristics.text: (1) Genetics and Genome Biology, SickKids Research Institute, Toronto, ON, Canada (2) Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada
characteristics.characteristic: INSDC center name
characteristics.text: 2023-04-01T00:23:18Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-04-01T00:23:18Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS13629691
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12340:WL1031
characteristics.characteristic: Submitter Id
characteristics.text: 32
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Male and female mice were euthanized using automated Euthanex Carbon Dioxide (CO2) lids followed by decapitation between 9:30 am and 11:30 am. Whole hypothalamus were dissected on postnatal days 12, 22, 27, 32 and 37. Collected tissues were directly placed in RNAlater (containing 10% w/v sodium citrate tribasic dihydrate and 60% w/v ammonium sulphate following dissection, and stored at −20 °C until RNA extraction. Total RNA was extracted from mouse hypothalamus with the NucleoSpin® miRNA kit (Macherey Nagel) in combination with TRIzol lysis (Invitrogen) following manufacturers' protocols, allowing for collection of small RNA (< 200 nt) and large RNA (> 200 nt) simultaneously into separate tubes from total RNA. Prior to RNA extraction, tissue samples were placed into bead mill tubes containing six 1.4 mm ceramic beads (MoBio Laboratories), and homogenized for 30 s at 6.5 m/s at 4 °C using Omni Bead Ruptor 24 bead mill. RNA quantity was determined using Nanodrop and the quality was assessed by Agilent 2100 Bioanalyzer. Construction of RNA-seq libraries was automated with QuantSeq 3'mRNA-seq Library Prep Kit (Lexogen GmbH, Vienna) using the Agilent NGS Workstation (Agilent Technologies, Santa Clara) as per manufacturer's protocol. 250 ng of RNA, from the large RNA fraction, was used to generate cDNA. cDNA was amplified with 17 PCR cycles as determined by qPCR analysis using the PCR Add-on kit (Lexogen). The resulting libraries were quantified with Qubit DNA HS (Thermo Fisher, Waltham) and fragment sizes analyzed on the Agilent Bioanalyzer using the High Sensitivity DNA assay prior to sequencing.
characteristics.characteristic: description
characteristics.text: peri-puberty
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: WL1031_Hd32F4
characteristics.characteristic: individual
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: hypothalamus
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: WL1031
characteristics.characteristic: title