characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-09-02T17:00:24Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-09-02T17:00:24Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA113597714
characteristics.characteristic: External Id
characteristics.text: Lund University
characteristics.characteristic: INSDC center name
characteristics.text: 2023-09-02T17:00:24Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-09-02T17:00:24Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS15591803
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-13060:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: 14
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: thale cress
characteristics.characteristic: common name
characteristics.text: Col-0
characteristics.characteristic: cultivar
characteristics.text: Protocols: Mitochondria were collected from two-week-old in vitro grown plants in shaking flasks. Plants were grown in soil-filled pots in long day conditions (16h light) at approximately 120 umol s-1 m-2. RNA immunoprecipitation (RIP) assays were performed with the μMACS GFP-Tagged Protein Isolation Kit (Miltenyi Biotec) according to manufacturer’s instructions with minor modifications. Samples were homogenized in RIP lysis buffer (20 mM HEPES-KOH, pH 7.6, 100 mM KCl, 20 mM MgCl2, 1 mM DTT, 1% (v/v) Triton X-100, 1X of complete EDTA-free protease inhibitor (Roche)) for 30 min at 4°C with slow rotation (10 rpm). The lysates were clarified by centrifugation at 100,000 g for 20 min at 4°C and resulting the supernatant was incubated with 50 μL of anti-GFP magnetic beads (Miltenyi Biotec) for 30 min at 4°C with rotation (10 rpm). Following three washes with RIP washing buffer (lysis buffer with 0.1% Triton X-100) and recovery of the beads on a magnetic stand each time, RNAs were extracted from the beads with TRIzol reagent (Life Technologies), precipitated with ethanol, and then used for RT-qPCR analysis. Prior to retrotranscription, immunoprecipitated RNAs were treated with DNase Max (QIAGEN). Ribosome footprints were depleted from ribosomal RNAs after first strand cDNA synthesis using custom biotinylated oligonucleotides matching major rRNA contaminants found in previous experiments (Planchard et al., 2018).N. Planchard, P. Bertin, M. Quadrado, C. Dargel-Graffin, I. Hatin, O. Namy, H. Mireau, The translational landscape of Arabidopsis mitochondria. Nucleic acids research 46, 6218-6228 (2018).
characteristics.characteristic: description
characteristics.text: LP.04 four leaves visible stage
characteristics.characteristic: developmental stage
characteristics.text: 35S::mito-GFP
characteristics.characteristic: genotype
characteristics.text: not collected
characteristics.characteristic: geographic location (country and/or sea)
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Arabidopsis thaliana
characteristics.characteristic: organism
characteristics.text: Arabidopsis thaliana
characteristics.characteristic: organism
characteristics.text: whole organism
characteristics.characteristic: organism part
characteristics.text: Arabidopsis thaliana
characteristics.characteristic: scientific_name
characteristics.text: none
characteristics.characteristic: stimulus
characteristics.text: Sample 2
characteristics.characteristic: title