HiSeq X Ten paired end sequencing; RNA-seq of oocytes and zygotes from pabpn1l ko and wt female mice
RNA-seq of oocytes and zygotes from pabpn1l ko and wt female mice
GV oocytes and zygotes were collected and mixed from three mice of each genotypes seperately. Each group contained a total of 30 oocytes or zygotes. In the initial step, we spiked 2 × 106 mRNA-EGFP, transcribed in vitro, into each group. Each cell sample after sorting can be stored in 6μl Discover-sc™ WTA Kit V2(Vazyme, N711, Nanjing, China) sample buffer at -70°C. The volume of fresh cells was measured, and Discover-sc™ WTA Kit V2 was used for cell lysis and first strand cDNA synthesis. Then, the first stand cDNA was amplified by Discover-sc PCR Mix. The amplified double stranded cDNA was purified by AMPure XP beads. The PCR products were purified by AMPure XP beads and the final library was obtained.