Blastocysts derived from young and aged father mice.

Paternal aging affects success in pregnancy and increases risk in their offspring. Paternal aging associated events in embryos will help understanding the aged associated low pregnancy out come and high risk in their offspring. This data is RNAseq of the blastocysts of young aged male mice. The same C57/BL6N mice were mate with female ICR mice (8-10 weeks) at their younger (13-23 weeks) and aged (50-55 weeks) lifetime. ICR female were super ovulated (7.5 IU equine chorionic gonadotropin followed by intraperitoneal administration of 7.5 IU human chorionic gonadotropin) and the mice were mate with male mouse. Twenty-four hours after hCG injection, 2-cell stage embryos were collected from oviducts and cultured for 2 days upto the blastocysts stage. For preparation of RNA of each experimental replicate, thirty-five blastocysts were used. RNA in embryos were extracted using RNAqueous (Invitrogen) and the RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA). Average RIN is 9.0. Libraries were created using NEBNext Single Cell/Low Input RNA Library Prep Kit and the library quality and quantity were determined using the Agilent 2100 Bioanalyzer and the KAPA Library Quantification Kit (KAPA Biosystems, Wilmington, MA, USA), respectively. Clusters were generated on a cBot (Illumina), and one lane of the multiplied samples was sequenced as 100 bp reads (single read) on the NextSeq1000 (Illumina). Image analysis, base calling were performed using the RTA version 2.4.11 and bcl2fastq2 v2.20.0.422 (Illumina) following the manufacturers instructions. Sequence data were filtered to discard the adapter sequence, ambiguous nucleotides and low-quality sequences. Average sequence reads were 30,502,835 for each sample. The remaining sequence data were aligned to the Mus Musculus genome sequence (GRCm38) to count sequence reads.