Transcriptome characterization of sperm sexed by flow cytometry or density gradient.

• Grant number: 11/16826-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2012
• End date: February 28, 2014
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Vera Fernanda Martins Hossepian de Lima
• Grantee: William Jardim de Oliveira Santos
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

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.

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SANTOS, William Jardim de Oliveira. Caracterização do transcriptoma e da produção embrionária de espermatozóides sexados por citometria de fluxo ou por gradiente de densidade. 2014. Master's Dissertation - Universidade Estadual Paulista (Unesp). Faculdade de Ciências Agrárias e Veterinárias. Jaboticabal Jaboticabal.