characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-24T00:21:31Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-24T00:21:31Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110460919
characteristics.characteristic: External Id
characteristics.text: EMBL
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-24T00:21:31Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-24T00:21:31Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12558947
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12068:SN011_bcd4x_evolved
characteristics.characteristic: Submitter Id
characteristics.text: 2h30min to 3h30min
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: fruit fly
characteristics.characteristic: common name
characteristics.text: Protocols: 2.5h-to-3.5h-old embryos (developed in room temperature) were dechorionated and flash-frozen in liquid nitrogen for nuclei preparation. The evolved embryos are from population 2-6-1A, at the 20th generation. They were manually examined and smaller embryos were removed upon collection, to reduce noise and focus on relatively large embryos. A wild-type line (VK33) and the 4xbcd lab stock were treated in parallel, however without manual sorting. Nuclei isolation was performed following a standard protocol (10x Genomics Single Cell Protocols, with adaptations from Francisca Hervas-Sotomayor at Heidelberg University). The frozen embryos were squished with a pestle for 20 times in cold homogenisation buffer (HB) [250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 10 mM Tris-HCl (pH 8), 0.1% Nonidet P40/IGEPAL, 1 uM DTT, 0.4 U/ul RNAse Inhibitor (New England Biolabs), 0.2 U/ul SUPERase In RNase Inhibitor (Invitrogen)]. The samples were then centrifuged at 100 g for 1 min to remove unlysed tissue, and the supernatant was centrifuged at 1,000 g for 5 min to pellet the nuclei. The pellet was washed once in HB, filtered twice with Flowmi Cell Strainers (Sigma) and resuspended in PBS. A subsample of the nuclei prep was DAPI-stained and examined under the microscope, to determine the density of nuclei. For each sample, 7,500 nuclei were used as the input for 10x library construction. The library was constructed following standard 10x Genomics protocol (Chromium Next GEM Single Cell 3_ Reagent Kits v3.1).
characteristics.characteristic: description
characteristics.text: Generation 20
characteristics.characteristic: generation
characteristics.text: transgenic
characteristics.characteristic: genetic modification
characteristics.text: bicoid-eGFP inserted at VK33
characteristics.characteristic: genotype
characteristics.text: egg laying
characteristics.characteristic: initial time point
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Drosophila melanogaster
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.characteristic: scientific_name
characteristics.text: mixed sex
characteristics.characteristic: sex
characteristics.text: VK33
characteristics.characteristic: strain
characteristics.text: SN011_bcd4x_evolved
characteristics.characteristic: title
characteristics.text: EMS-mutagenesis at G0
characteristics.characteristic: treatment