2025-10-30
ENA first public
ERC000011
ENA-CHECKLIST
Research Institute for Farm Animal Biology (FBN) Physiological Genomics Unit
INSDC center name
public
INSDC status
ERS26815324
SRA accession
E-MTAB-15588:L169
Submitter Id
day 63 of gestation
day
age
ArrayExpress
broker name
not collected
collection date
pig
common name
Protocols: Briefly, twenty-two F₁ sows from (German Landrace [GL] × Piétrain [Pi]) were backcrossed to three GL and three Pi sires to produce F₁×GL and F₁×Pi conceptuses. A total of 270 BC fetuses were collected at day 63 of gestation. From these, 38 fetuses were selected for this study. Tissues (liver, brain, muscle, kidney, ovary, and testis) were carefully dissected, immediately snap-frozen in liquid nitrogen, and stored at −80°C until RNA extraction. For the four non-gonadal tissues (liver, brain, muscle, and kidney), a set of 24 matched samples was obtained from the same fetuses (n=12 per BC group for each tissue). Furthermore, 17 ovary samples (9 F₁ × GL, 8 F₁×Pi) and 18 testis samples (10 F₁ × GL, 8 F₁×Pi) were collected. Of these gonadal samples, 10 ovaries and 11 testes originated from the same 24 fetuses used for the non-gonadal tissues. The remaining 7 ovary and 7 testis samples were obtained from additional fetuses not included in the non-gonadal set.  Total RNA was isolated from tissue samples using QIAzol Lysis Reagent (Qiagen, Germany), while the ovary, testis, and muscle tissues underwent additional homogenization with ceramic beads using a Precellys 5000 homogenizer. These tissues were chosen for homogenization as they are denser and more fibrous, requiring additional mechanical disruption for effective RNA extraction. Following RNA extraction, DNase treatment was applied to remove genomic DNA contamination, and RNA was purified using the RNeasy Mini Kit (Qiagen, Germany). RNA concentration and quality were assessed using a NanoDrop ND-1000 spectrophotometer (Peqlab) and a Bioanalyzer 2100 (Agilent Technologies). To prepare the RNA library, 1 µg of total RNA with a RIN > 8 was used for library preparation using an Illumina Stranded mRNA Prep, Ligation kit with 11 PCR-cycles of amplification according to the manufacture's recommendation (Illumina). The libraries were quality checked for fragment length distribution on Agilent 2100 Bionalyzer and normalized to an equal concentration of 10nM prior to pooling.
description
fetus
developmental stage
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Sus scrofa
organism
Sus scrofa
organism
Liver
organism part
Sus scrofa
scientific_name
male
sex
F1 (German Landrace × Pietrain) X German Landrace
strain
L169
title