Micro RNA in culture medium of bovine oocyte granulosa cells complexes

MicroRNA is a potential regulator of oocyte development and accumulated in follicular fluids. Follicular fluid is a sole environment for oocyte growth, and miRNAs in follicular fluids have been proved to play a role in oocyte growth. This sample is miRNAs extracted from the culture medium of oocyte granulosa cells complexes (OGCs). OGCs were collected from early antral follicles (0.5-0.7mm in diameter) of cows and cultured for 14 days. Two days before the sampling, the medium was exchanged with alpha-MEM medium containing exosome depleted serum (EXO-FBS-50A, SBI). At day 16 of culture period, the medium was collected and exosome RNA in the culture medium were extracted using a SeraMir Exosome RNA Amplification Kit (System Biosciences). RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA); a cDNA library was constructed using an Illumina TruSeq Small RNA Library Preparation Kit (Illumina, San Diego, CA, USA) for RNA from FFs. Clusters were generated on a cBot (Illumina), and one lane of the multiplied samples was sequenced as 75 bp reads (single read) on the NextSeq (Illumina). Image analysis, base calling and quality filtering were performed using the bcl2fastq2 v2.18.0.12 (Illumina) following the manufacturer's instructions. Sequence data were filtered to discard the adapter sequence, ambiguous nucleotides and low-quality sequences. The remaining sequence data were aligned to the Bos Taurus genome sequence (ARS-UCD1.2/bosTau9) to count sequence reads.