characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-12-08T14:39:03Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-12-08T14:39:03Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA10159449
characteristics: External Id
characteristics: Southern University of Science And Technology
characteristics: INSDC center name
characteristics: 2022-12-08T14:39:03Z
characteristics: INSDC first public
characteristics: 2022-12-08T14:39:03Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS7815934
characteristics: SRA accession
characteristics: E-MTAB-10966:H3K27me3_Seedling_IP_rep3
characteristics: Submitter Id
characteristics: ArrayExpress
characteristics: broker name
characteristics: thale cress
characteristics: common name
characteristics: Col-0
characteristics: cultivar
characteristics: Protocols: Open flower buds were freshly collected from pollen marker line into 9% sucrose solution and matured pollen grain were enriched by vortex and filtered through a 60 um nylon mesh. The sperm nuclei and vegetative nuclei were isolated by FACS using a method described previously (cite paper). Briefly, pollen grain were resuspended in ice-cold Galbraith buffer (45mM MgCl2, 30mM Sodium Citrate, 20mM MOPS, 1% Triton X-100, pH to 7.0) supplemented with 1X complete protease inhibitor cocktail (Roche) and pollen wall was then disrupted by additional vortexed at 60Hz for 30sec with glass beads (sigma). The resulting pollen wall debris were removed by filtering through a 10 um nylon mesh. The crude suspension was ready for FACS on a FACSAria III cell sorter. FACS was carried out in standard configuration using 70um nozzle. A 488nm laser was used for GFP excitation and a 561nm laser for RFP excitation. The pollen marker line (ACT::H2B-mRFP labels vegetative nucleus, MGH3P::MGH3-eGFP labels sperm nucleus in mature Arabidopsis pollen) used in this study was kindly provided by prof. Jorg Dieter Becker (Santos et al. 2017 doi.org/10.1007/978-1-4939-7286-9_16). Plants were grown in climate controlled greenhouse under long day conditions (16hr light, 8hr dark) at 22C. ChIPed DNA was purified using Qiagen MinElute PCR Purification Kit. Library DNA was purified using AmpureXP beads using 1:1 ratio. Antibodies used: anti-H3k27me3 (Millipore 07-449); and anti-H3K4me3 (Active Motif 39915) and Libraries were constructed using Nextera DNA Preparation Kit.
characteristics: description
characteristics: seedling development stage
characteristics: developmental stage
characteristics: wild type genotype
characteristics: genotype
characteristics: Arabidopsis thaliana
characteristics: organism
characteristics: Arabidopsis thaliana
characteristics: organism
characteristics: whole plant
characteristics: organism part
characteristics: Arabidopsis thaliana
characteristics: scientific_name
characteristics: H3K27me3_Seedling_IP_rep3
characteristics: title