characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-05-30T17:01:53Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-05-16T18:32:13Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104038019
characteristics.characteristic: External Id
characteristics.text: unspecified
characteristics.characteristic: INSDC center name
characteristics.text: 2017-05-30T17:01:53Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-05-16T18:32:13Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1701912
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5730:Control-01
characteristics.characteristic: Submitter Id
characteristics.text: 2
characteristics.unit: month
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Upon isofluorane anesthetization, two months-old transgenic C57BL/6 female mice of around 20-25 gr were locally injected with α-LTx close to the LAL muscles. Animals were randomized in the experimental groups based on their body weight. The toxin (5 μg/kg) was diluted in 15 μl of physiological saline (0.9% w/v NaCl in distilled water). Control animals were injected with saline. At different time points (0, 4, 24 and 96 h) treated mice were sacrificed by anesthetic overdose followed by cervical dislocation and LAL muscles were immediately fixed with a local injection PFA 4% for 10 min. Muscles were collected, incubated with fluorescent α-BTx in sterile PBS for 30 10 min at 37°C and then frozen in liquid nitrogen. Cryo-sections (7 μm thick) were transferred to UV treated microscope glass slides. Microdissection was performed under direct microscopic visualization Total RNA was isolated from the microdissected samples by incubation with 50 μl of lysis buffer PKD (Qiagen, Venlo, Netherlands) and 10 μl of proteinase K solution (Promega, Madison, WI, USA) at 55 °C over night with the sample upside down. The day after samples were centrifuged for 10 min at 10000 rpm and RNA extracted using the Maxwell® 16 LEV RNA FFPE Purification Kit (Promega, Madison, WI, USA) following manufacturer’s instructions. To prepare cDNA from RNA samples, the SMARTer Universal Low Input RNA kit (Clontech Laboratories) was employed following manufacturer’s instructions Libraries were prepared using the Ion TargetSeq Exome Enrichment kit, following the manufacturer’s guidelines.
characteristics.characteristic: description
characteristics.text: Plp1-EGFP
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: neuromuscular junction
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/GO_0031594
characteristics.characteristic: organism part
characteristics.text: levator auris longus muscle
characteristics.characteristic: sampling site
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: C57BL/6
characteristics.characteristic: strain
characteristics.text: Control-01
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001461
characteristics.characteristic: title