title: Neurite_shLarp1_2
organism: Mus musculus
description: Protocols: Primary cortical neurons were prepared from P0 mouse pups (C57B/6), and cultured as previously described (Kaech, 2006). 106 primary neurons were plated per a Millicell cell culture insert (6-well, Millipore MCRP06H48), coated with poly-D-Lysine and laminin. Infection with lentiviral particles was at DIV5 and cells were harvested 72 hours later (DIV9). Separation into soma and neurites was performed using a cotton swab as described (Zappulo, 2017) and further separation of soma on nuclear and cytoplasmic fractions was performed using NE-PER Nuclear and Cytoplasmic Extraction Kit (Thermo 78833) following manufacturer's instructions. RNA was extracted from Trifast using 2-step chloroform extraction and isopropanol precipitation. RNA samples were quantified using qubit and analysed for integrity with tapestation. Good separation was verified by RT-qPCR. TrueSeq stranded mRNA Library kit; 1ul of 1:300 ERCC1 spike-in was added to 49ul initial solution with 150ng total RNA.; Final PCR was done with 12-16 cycles' reaction
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-07-12T00:19:24Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-07-12T00:19:24Z
attributes.tag: External Id
attributes.value: SAMEA113399762
attributes.tag: INSDC center alias
attributes.value: Berlin Institute for Medical Systems Biology, Max Delbruck Center for Molecular Medicine, Berlin, Germany
attributes.tag: INSDC center name
attributes.value: Berlin Institute for Medical Systems Biology, Max Delbruck Center for Molecular Medicine, Berlin, Germany
attributes.tag: INSDC first public
attributes.value: 2023-07-12T00:19:24Z
attributes.tag: INSDC last update
attributes.value: 2023-07-12T00:19:24Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12886:Neurite_shLarp1_2
attributes.tag: age
attributes.value: 0
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: neuron of crebral cortex
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: neonate
attributes.tag: fraction
attributes.value: neurite
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: brain
attributes.tag: sample name
attributes.value: E-MTAB-12886:Neurite_shLarp1_2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: mixed
attributes.tag: strain
attributes.value: C57BL/6