characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-09-25T00:22:23Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-09-25T00:22:23Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA111393317
characteristics: External Id
characteristics: European Research Institute for the Biology of Ageing, University Groningen, UMCG Groningen, The Netherlands
characteristics: INSDC center name
characteristics: 2022-09-25T00:22:23Z
characteristics: INSDC first public
characteristics: 2022-09-25T00:22:23Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS13490295
characteristics: SRA accession
characteristics: E-MTAB-7399:P0_Rep2
characteristics: Submitter Id
characteristics: ArrayExpress
characteristics: broker name
characteristics: cerebellar granule neuron progenitor
characteristics: cell_type
characteristics: house mouse
characteristics: common name
characteristics: Protocols: Mice were conventionally housed, fed ad libitum, and routinely genotyped by PCR. Timed matings were performed overnight, with the following morning considered E0.5. Pregnancies were detected by measuring female weight gain at E13.5. A subset of pregnant females received a single dose of Tamoxifen (2 mg/100 μl peanut oil, Sigma) by oral gavaging at E13.5. Pregnant females were killed by asphyxiation (CO2), neonatal mice until the age of P7 were killed by decapitation. For transcriptional analyses, CGNPs were harvested from E13.5, E15.5, P0, P7, P14, and P30 tdTomato+ dissected cerebella from offspring from Tamoxifen-treated females. For primary cell cultures, CGNPs were harvested from E13.5, E15.5 and P7 days old cerebella from offspring of non-treated females. Cerebellar dissection was performed using a stereomicroscope. E15-P30 Cerebella were dissociated with a Papain dissociation kit following the manufacturer’s instructions (Worhtington). Following papain treatment, ovomucoid was added to stop the reaction. For P14 and P30 cerebella, an additional Percoll gradient step was performed to remove myelin and debris. Cell suspensions were filtered through a 70 μm cell strainer prior to further processing. Cerebellar single cell suspensions (E15.5-P30) were subjected to fluorescenceactivated cell sorting (FACS) for tdTomato+ cells, using a MoFlo Astrios cell sorter (Beckman Coulter). tdTomato+ cells (CGNPs) were collected in 1.5 ml tubes, centrifuged, and snap frozen. RNA was extracted using the NucleoSpin RNA XS kit (Macherey-Nagel), following the manufacturer's instructions. RNA concentration was measured with a QuBit RNA HS assay kit (Thermo Fisher) and analyzed on a bioanalyzer using the Agilent RNA 6000 Pico Kit (Agilent). 2,5-10 ng of RNA per sample was used as input for library preparation. For library prepping, the Clontecht SMARTer stranded total RNA-seq kit-pico was used according to the manufacturer’s instructions for mammalian sample input.
characteristics: description
characteristics: P0
characteristics: developmental stage
characteristics: Math1-CreERT2 tdTomato+
characteristics: genotype
characteristics: A8
characteristics: individual
characteristics: Mus musculus
characteristics: organism
characteristics: Mus musculus
characteristics: organism
characteristics: cerebellum
characteristics: organism part
characteristics: Mus musculus
characteristics: scientific_name
characteristics: not available
characteristics: sex
characteristics: Math1-CreER[T2]
characteristics: strain
characteristics: P0_Rep2
characteristics: title