Instrument: Illumina HiSeq 1500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Oocytes were freshly collected as described in Verlhac et al, Development 120, 1017-1025 (1994). Total RNA was extracted from oocytes using the RNAqueous®-Micro Total RNA Isolation Kit (Thermofisher, Ref. AM1931). Oocytes were washed 3 times in PBS, resuspended in lysis buffer, freezed in liquid nitrogen and conserved at -80°C overnight. After unfreezing, RNA extraction was carried out following the manufacturer’s instructions. Library preparation and Illumina sequencing were performed at the Ecole normale superieure Genomic Platform (Paris, France). 1 ng of total RNA were amplified and converted to cDNA using NuGEN’s Ovation RNA-Seq kit V2. Following amplification, 1 µg of cDNA was fragmented to ∼200 bps using Covaris S200. The remainder of the library preparation was done using 200 ng of cDNA following TruSeq RNA Sample Prep v2 kit from the End Repair step. Three runs were performed following the same design: libraries were multiplexed by 4 on 1 flow cell lane. A 50 bp read sequencing was performed on a HiSeq 1500 device (Illumina). A mean of 17,3 ± 3,9 million passing Illumina quality filter reads was obtained for each of the 4 samples.