title: GSM5008083: PIWIL1hetero - RNA-seq_rep3; Mesocricetus auratus; RNA-Seq
library_construction_protocol: Mature females were induced to superovulate by i.p. injection of 40 IU PMSG (ASKA Animal Health) at 1200 to 1400 on the day of conspicuous, postestrus vaginal discharge (day 1 of the estrous cycle). GV oocytes were collected from ovary after 102h PMSG injection. MII oocytes were collected from oviduct after 115h PMSG injection. PN zygotes were collected from oviduct after 91 or 115h PMSG injection followed by mating with male during the night of day4 or 5 (depend on the female estrus cycle). small-RNA-seq: Total RNAs for small RNA sequence were extracted from MII oocytes of PIWIL1 and PIWIL3 homozygous and heterozygous mutants. The construction of small RNA libraries was performed using NEXTFLEX® Small RNA-Seq Kit v3 (PerkinElmer) with the manufacturer's instructions. RNA-seq: Total RNA for RNA-seq were purified from 50 MII oocytes of 2 to 4-month-old golden hamsters using NucleoSpin RNA Plus XS (MACHEREY-NAGEL). RNA-seq libraries were construct using total RNA from MII oocytes and SMART-Seq Stranded Kit (TaKaRa Bio). Bisulfite-seq: DNA from thirty GV oocytes were spiked with 1% unmethylated lambda phage DNA. Libraries were generated by post bisulfite adaptor tagging method with 8 cycles of library amplification.