2025-11-12
ENA first public
ERC000011
ENA-CHECKLIST
Institute of Molecular Biology
INSDC center name
public
INSDC status
ERS27068435
SRA accession
E-MTAB-15730:wt_25_rep2
Submitter Id
embryo
dimensionless unit
age
ArrayExpress
broker name
not collected
collection date
Protocols: Synchronized gravid adults were subjected to hypochlorite treatment to isolate embryos. Embryos were washed four times with cold M9 buffer and snap-frozen as \"worm balls\" in liquid nitrogen.  C. elegans strains were maintained at 20°C on NGM plates seeded with E. coli OP50. Animals were synchronized by hypochlorite treatment and hatched in M9 buffer for 16 hours. Synchronized L1 larvae were transferred to either 15°C or 25°C incubators and grown to gravid adult stage. Embryos were harvested by hypochlorite treatment when the first laid eggs were observed on plates, approximately 3-4 days post-L1 transfer depending on temperature. Frozen samples were ground with mortar and pestle, then mixed with five volumes of TRIzol™ LS Reagent (Invitrogen™). RNA was extracted using Direct-zol RNA Microprep kit (Zymo Research™) with additional TURBO DNase treatment (Invitrogen™) to remove genomic DNA. RNA was resuspended in nuclease-free water. NGS libraries were prepared using Illumina's Stranded mRNA Prep Ligation Kit with 1000 ng starting material and 10 amplification cycles. ERCC spike-ins (2 μl of 1:100 dilution, Ambion) were added to assess technical variability and optionally assist with library size normalization. Libraries were profiled using DNA 1000 chip on 2100 Bioanalyzer (Agilent) and quantified with Qubit dsDNA HS Assay Kit.
description
C. elegans embryo stage
developmental stage
wild type genotype
genotype
not collected
geographic location (country and/or sea)
25°C
growth condition
11
individual
not applicable
isolate
Caenorhabditis elegans
organism
Caenorhabditis elegans
organism
whole organism
organism part
Caenorhabditis elegans
scientific_name
hermaphrodite
sex
N2
strain
wt_25_rep2
title