RNA-Seq of pig healthy endometrium: sample 1 of day 28 of pregnancy
The library was prepared by an rRNA depletion method. Briefly, the ribosomal RNA was depleted from total RNA using the Epicentre Ribozero rRNA Removal Kit (Epicenter, USA) following the manufacturers protocol. Subsequently, the RNA was fragmented into 250~300 bp, and the fragmented RNA and dNTPs (dATP, dTTP, dCTP, and dGTP) were used for reverse transcription of the first-strand cDNA. RNA was degraded using RNase H, and second-strand cDNA was synthesized using DNA polymerase I and dNTPs. Then, through the exonuclease/polymerase activities, the remaining overhangs of double-stranded cDNA were converted to blunt ends. After the 3' ends of the DNA fragments were adenylated, sequencing adaptors were ligated to the cDNA. The library was purified by using the AMPure XP system to select cDNA fragments preferably 350-400 bp. Uridine digestion was performed using Uracil-N-Glycosylase, which was followed by the cDNA amplification using PCR.