biostudies-arrayexpressJasmina Al-MousawiMus musculushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15453RNA-seq of single Fully Grown Oocytes using SMART-seq2. The oocytes were collected from female FVB mice, approximately. 44-48 h post-PMSG injection. The oocytes were microinjected with 1.3-1.7 μg/μL Kdm5b wild-type (WT) or catalytic mutant (CM) mRNA. Noninjected oocytes are denoted as “Noninj”. The oocytes were cultured overnight for approximately 16 h in M2 containing 20 μM Milrinone and 5% FBS and collected for sequencing.biostudies-arrayexpressGrowth Protocol - Ovaries were isolated from female mice 44-48 h post-PMSG injection. Cumulus-oocyte-complexes were released from the ovaries by mechanical disruption with a needle into M2 supplemented with 20 μM Milrinone to prevent meiotic progression to the MII phase. Cumulus cells were manually removed from the fully grown oocytes.Nucleic Acid Extraction - The RNA was reverse transcribed to generate cDNA and purified with a 0.6 x ratio of SPRIselect beads according to previously published protocols (Picelli et. al. 2014, Hennig et. al. 2018).Sequencing - The libraries were pooled and sequenced on a NextSeq2000 Sequencer using 50 bp read lengths in paired-end mode. The individual libraries were sequenced to an average depth of 5 million reads per sample.Library Construction - SMART-seq2 libraries were prepared by the EMBL Genecore Facility, as previously described protocols (Picelli et. al. 2014, Hennig et. al. 2018).Sample Treatment - Microinjections were performed immediately after the collection of fully grown oocytes (FGOs) using an Eppendorf Femtojet system with a Leica iDM8 microscope. FGOs were washed in warm M2 media and placed in an M2 drop under mineral oil. Kdm5b WT and CM mRNAs were microinjected into the cytoplasm of FGOs at 1.3-1.7 μg/μL. After injection, FGOs were cultured in M2 containing 20 μM Milrinone and 5% FBS under 20 % O₂ and 5% CO₂ at 37°C for 14-16 h before collection.Sample Collection - At the appropriate stage, oocytes were washed in 37°C M2. Oocytes were collected individually using a mouth pipette and lysed. Samples were stored at -20°C until library preparation.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - Fastq files were mapped to the GRCm38/mm10 genome containing only major chromosomes (chr1-18, X, Y and MT) using STAR. For SMART-seq2 data generated in this study, paired-end reads were mapped using alignreads mode with the following parameters: --outFilterMultimapNmax 20 \ --outFilterMultimapScoreRange 0 \ --outFilterMismatchNoverLmax 0.05 \ --sjdbScore 2. Read quantification was performed using Featurecounts and options -p --countReadPairs.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsNextSeq 2000RNA-seq of coding RNAMus musculusVladimir BenesLaura VillacortaJasmina Al-MousawiAna BoskovicfalseSMART-seq2 of Kdm5b Overexpression in Fully Grown OocytesRNA-seq of single Fully Grown Oocytes using SMART-seq2. The oocytes were collected from female FVB mice, approximately. 44-48 h post-PMSG injection. The oocytes were microinjected with 1.3-1.7 μg/μL Kdm5b wild-type (WT) or catalytic mutant (CM) mRNA. Noninjected oocytes are denoted as “Noninj”. The oocytes were cultured overnight for approximately 16 h in M2 containing 20 μM Milrinone and 5% FBS and collected for sequencing.2026-06-24T00:00:00Z2026-06-24T14:20:04.478Z2025-08-05T21:05:39.603ZE-MTAB-15453ERP177857EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003816EFO_0003738EFO_0003969EFO_0004184