The aim of this proposal is to characterize the transcriptome of sperm from sexed by density gradient centrifugation and flow cytometry. The experimental groups for semen and embryos going to be: 1) fresh semen, 2) control group: conventional semen thawed, 3) group gradient: density gradient centrifugation 3) group cytometry: sexed semen by flow cytometry. To obtain data on gene expression in sperm, identification of SNPs (single nucleotide polymorphisms) and alternative splicing will be built libraries Paired Ends (2x100bp), covering 100 million reads per sample. The sequencing reactions going to be performed in high-performance platform IIx Genome Analyzer (Illumina, San Diego, CA, USA). The sequencing reactions going to be performed in the transcriptome platform SOLiD " 3 System (Applied Biosystems). Genes differentially expressed between the pools will be validated by real-time PCR (qPCR), using the samples included in each pool separately.
News published in Agência FAPESP Newsletter about the scholarship: