GSM6171998: Four weeks Fasted pigID=12; Sus scrofa; RNA-Seq
Piglets were anasthesized using a mixture of Zoletil+Xylazine (5:2 ratio, 1 mg/10 kg BW), and euthanized by lethal injection with pentobarbital (24 mg/kg BW), followed by exanguination. The intestine was isolated, and mid-jejunum was identified at 50% of the small intestinal length. Intestine was washed in cold PBS, dabbed dry and mucosa was scraped off using clean glass slides. Collected tissue was snap frozen in liquid nitrogen and stored at -80°C until further analysis. Total RNA was extracted by grinding frozen muscle tissue and dissolving approximately 5-10 mg in RLT buffer containing b-mercaptoethanol. Further RNA isolation was performed according to the RNAeasy mini kit instructions (Qiagen). RNA concentration was measured using NanoDrop spectrophotometer (Thermofisher). RNA integrity was measured using RNA Screentape on the 2200 Tapestation (Agilent). RNA preparation, library construction and sequencing on BGISEQ-500 was performed at Beijing Genomics Institute.