SN011_bcd4x_evolved
Drosophila melanogaster
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).
ENA-FIRST-PUBLIC
2023-07-24T00:21:31Z
ENA-LAST-UPDATE
2023-07-24T00:21:31Z
External Id
SAMEA110460919
INSDC center alias
EMBL
INSDC center name
European Molecular Biology Laboratory
INSDC first public
2023-07-24T00:21:31Z
INSDC last update
2023-07-24T00:21:31Z
INSDC status
public
Submitter Id
E-MTAB-12068:SN011_bcd4x_evolved
age
2h30min to 3h30min
broker name
ArrayExpress
common name
fruit fly
generation
Generation 20
genetic modification
transgenic
genotype
bicoid-eGFP inserted at VK33
initial time point
egg laying
isolate
not applicable
sample name
E-MTAB-12068:SN011_bcd4x_evolved
scientific_name
Drosophila melanogaster
sex
mixed sex
strain
VK33
treatment
EMS-mutagenesis at G0