title: Illumina Genome Analyzer IIx sequencing; RNA-seq of estrogen (E2) induced changes of mRNA expression in early pregnant porcine endometrium
design: RNA-seq of estrogen (E2) induced changes of mRNA expression in early pregnant porcine endometrium
library_name: Sample 8_s
library_construction_protocol: The uterus was flushed with 10 ml and another 50 ml of phosphate-buffered saline (autoclaved, ph 7.4). Then, endometrial samples were collected, shock frozen in liquid nitrogen and stored at – 80 °C.  We analyzed the effects of gestational estradiol-17β (E2) exposure on the endometrium. Two distinct low doses corresponding to the acceptable daily intake (ADI) and close to the no-observed effect level (NOEL) and a high dose (0.05, 10 and 1000 µg E2/kg body weight daily, respectively) were orally applied to sows from insemination until sampling at day 10 of pregnancy and compared to carrier-treated controls.  RNA was harvested by means of TRIzol reagent (n = 4 per group, except for the ADI group, where a repeated tissue extraction was done because of only n=3 available animals). The library preparation starting from 125 ng total RNA of each endometrial sample was performed with the Encore Complete RNA-Seq Multiplex System IB (NuGEN, AC Leek, The Netherlands) following the manufacturers' protocol.
attributes.tag: Experimental Factor: compound
attributes.value: 17beta-estradiol
attributes.tag: Experimental Factor: dose
attributes.value: 0.05