GSM9010131: Restricted-fed sow 9; Sus scrofa; RNA-Seq
Total RNA of the mural granulosa cells (GCs) from each sow was extracted using the TRIzol method following the manufacturer's instructions. The RNA Integrity Number (RIN) was assessed using the Agilent 2100 Bioanalyzer. In this study, the mean RIN was 7.30 ± 0.60 (± SD), and none of the RNA samples used in this study had a RIN below 5. One sample of the FF group, FF09, was excluded based on its low RIN number, and therefore subsequqently not taken along. rRNAs were removed from total RNA, followed by fragmentation (to 200~500 nucleotides). The first-strand cDNA was synthesized using a random hexamer-primer, and dTTP was substituted by dUTP during the synthesis of the second strand. Next, the short fragments were purified and treated with elution buffer to facilitate end repair, followed by the addition of a single nucleotide A (adenine). After that, the adapters were connected to the fragments, and UNG (uracil-N-glycosylase) was used to degrade the second strand. After agarose gel electrophoresis, the suitable fragments were selected for several rounds of PCR amplification.