2025-11-12
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Molecular Biology, Mainz
INSDC center name
public
INSDC status
ERS27074103
SRA accession
E-MTAB-15751:Strain10_100c
Submitter Id
adult
dimensionless unit
age
ArrayExpress
broker name
not collected
collection date
Protocols: After confirmation of suppressor phenotypes and backcrossing to the parental tost-1(xf196 ts) strain, suppressor strains were expanded for genomic DNA extraction. Mixed-stage worm populations were cultured on 90 mm NGM plates seeded with E. coli OP50 for one generation, then transferred to 150 mm plates for an additional generation to increase biomass. Worms were harvested by washing plates with M9 buffer, collected by centrifugation, and washed multiple times to remove bacteria. Worm pellets were processed immediately for DNA extraction using the Gentra Puregene Tissue Kit (Qiagen) following the manufacturer's protocol for animal tissues. DNA concentration and purity were assessed by spectrophotometry before proceeding to library preparation. Genomic DNA was extracted using Gentra Puregene Tissue Kit (Qiagen). DNA (1.5 μg) was diluted to 55 μl in TE buffer and fragmented using Covaris S2 sonicator (Intensity: 5; Duty Cycle: 10%; Cycles/burst: 200; Time: 120 s; 2 cycles). Fragmented DNA was analysed on TapeStation with High Sensitivity D1000 ScreenTape.  Libraries were prepared using NEBNext Ultra II DNA Library Prep Kit starting with 10 ng fragmented DNA and 9 PCR cycles. Libraries were profiled on 2100 Bioanalyzer (Agilent) and quantified using Qubit 1× dsDNA HS Assay Kit. All 22 samples were pooled equimolarly and sequenced on NextSeq2000 P2 FC (2×100 cycles plus 2×8 index cycles).
description
adult
developmental stage
to be defined10
genotype
not collected
geographic location (country and/or sea)
10
individual
not applicable
isolate
Caenorhabditis elegans
organism
Caenorhabditis elegans
organism
whole animal
organism part
Caenorhabditis elegans
scientific_name
hermaphrodite
sex
RFK1926
strain
Strain10_100c
title