characteristics.text: Sort VN and sperm to 3 fractions by clover + PI
characteristics.characteristic: Experiment setup
characteristics.text: SAMN32329662
characteristics.characteristic: External Id
characteristics.text: Gregor Mendel Institue (GMI)
characteristics.characteristic: INSDC center name
characteristics.text: 2023-02-22T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2024-06-09T13:51:37.439Z
characteristics.characteristic: INSDC last update
characteristics.text: SRS16220746
characteristics.characteristic: INSDC secondary accession
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Plant
characteristics.characteristic: NCBI submission model
characteristics.text: Plant.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: 1
characteristics.characteristic: Replicate
characteristics.text: SRS16220746
characteristics.characteristic: SRA accession
characteristics.text: sperm_low
characteristics.characteristic: cell_type
characteristics.text: Col-0 HTR10-Clover Dry pollen was harvested using the vacuum suction method (Johnson-Brousseau and McCormick, 2004), using 150μm, 60μm and 10μm mesh filters and then flash freezed and kept in -80°C. Sperm and vegetative nuclei were isolated by FACS following the previously described method (Borges et al., 2012). Collected pollen was hydrated by resuspension in an ice-cold Galbraith buffer (45 mM MgCl2, 30 mM Tri-Sodium Citrate, 20mM MOPS pH 7.0, 0.1% Triton X-100) with added 10mM ß-mercaptoethanol and 1× complete protease inhibitor cocktail (Roche)(Galbraith et al., 1983). Pollen suspension was then centrifuged for 1 minute at 10,000 g, 50 μl of 100 μm glass beads were added and vortexed for 4 minutes to disrupt pollen walls. The suspension was filtered through a 10 μm nylon mesh to retrieve sperm and vegetative nuclei. HTR10-Clover nuclei were then stained with propidium iodide (P4170, Sigma-Aldreich). Sperm and VN were sorted according to the Clover signal and propidium iodide (P4170, Sigma-Aldreich), sperm nuclei were further separated in the FACS to three approximately equal groups based on their propidium iodide fluorescence. This is the low PI fraction of the sperm from repeat 1 of this experiment.
characteristics.characteristic: description
characteristics.text: Flowering
characteristics.characteristic: dev_stage
characteristics.text: Col-0
characteristics.characteristic: ecotype
characteristics.text: HTR10-Clover
characteristics.characteristic: genotype
characteristics.text: USA
characteristics.characteristic: geo_loc_name
characteristics.text: Arabidopsis thaliana
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_3702
characteristics.characteristic: organism
characteristics.text: pollen
characteristics.characteristic: tissue
characteristics.text: Plant sample from Arabidopsis thaliana
characteristics.characteristic: title