organism: Sus scrofa
description: Protocols: We used German Landrace sows, which were inseminated with Pietrain sperm. At day 10 of pregnancy, the uterus was flushed with 10 ml and another 50 ml of phosphate-buffered saline (autoclaved, ph 7.4). All embryos were transferred into a petri-dish containing PBS. Single embryos were shock frozen in liquid nitrogen and stored at -80 °C. A low dose close to the no-observed effect level (NOEL) and a high dose (10 and 1000 µg E2/kg body weight daily, respectively) were orally applied to sows from insemination until sampling at day 10 of pregnancy. They were compared to carrier-treated controls. Total RNA from single embryos was extracted using the AllPrep RNA/DNA Micro Kit (Qiagen, Hilden, Germany) following the manufacturers protocol for cells with slight modifications. In brief, 700 µl of Buffer RLT Plus supplemented with 1% β-mercaptoethanol was added to the frozen embryo. Disruption was achieved by pipetting up and down and by a single brief vortexing. Homogenization was performed using a syringe and needle. After centrifugation of the lysate using a DNA spin column, the flow-through was processed following the protocol for “purification of total RNA containing small RNAs from cells”. In order to improve RNA purity, the column was incubated with Buffer RPE at step D3 and D4 before centrifugation for 4 min and 2 min, respectively. RNA elution was repeated using the first eluate to increase the final concentration. Library preparation was performed using the TruSeq Stranded mRNA Sample Prep Kit (Illumina, Inc, California, USA) according to the manufacturer's protocol.
attributes.tag: ENA first public
attributes.value: 2018-10-16
attributes.tag: ENA last update
attributes.value: 2017-11-30
attributes.tag: External Id
attributes.value: SAMEA104410644
attributes.tag: INSDC center alias
attributes.value: ETH Zurich, Animal Physiology, Institute of Agricultural Sciences, Zurich, Switzerland
attributes.tag: INSDC center name
attributes.value: ETH Zurich, Animal Physiology, Institute of Agricultural Sciences, Zurich, Switzerland
attributes.tag: INSDC first public
attributes.value: 2018-10-16T17:02:44Z
attributes.tag: INSDC last update
attributes.value: 2017-11-30T12:02:56Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6263:Sample 23
attributes.tag: breed
attributes.value: German Lambrace x Pietrain
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: pig
attributes.tag: developmental stage
attributes.value: embryonic day 10.5
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: blastocyst
attributes.tag: sample name
attributes.value: E-MTAB-6263:Sample 23
attributes.tag: sex
attributes.value: male