title: GSM1922681: AOF1_WT_2 (RNA-Seq); Mus musculus; RNA-Seq
library_construction_protocol: RNA-Seq To obtain MII oocytes, 6-8-week-old mice were injected intraperitoneally with 5 IU of pregnant mare’s serum gonadotrophin (PMSG, Sigma) and, 48 hours later, with 5 IU of human chorionic gonadotrophin (hCG, Sigma). Superovulated mice were euthanized 16 hours after hCG injection, and oocytes collected from the oviducts and released into a hyaluronidase/M2 solution for removal of cumulus cells. For RNA-Seq , three biological replicates were prepared for each genotype. Each sample, which contained 100 GV oocytes collected from 2-3 mice, was lysed directly in 1 μl of prelude direct lysis buffer (Nugen). RNA was then subject to amplification using the ovation RNA-seq system v2 (Nugen). Amplified cDNA was fragmented using Covaris and paired-end libraries were then constructed using the TruSeq RNA Sample Preparation Kit v2 (Illumina) starting from end repair.
attributes.tag: GEO Accession
attributes.value: GSM1922681