SRA029132 submitted by University of Alberta on 2011-01-24
Discovery of genes expressed by preimplantation porcine embryos using 454 pyrosequencing
In order to maximize sequence diversity, we first used the Ribo-SPIA* technology (NuGEN) to generate a cDNA library for 454 sequencing from total RNA extracted from pools of day 5-6 porcine blastocysts. Subsequently, we also constructed two normalized cDNA libraries from in vitro and in vivo generated embryos and subjected them to 454 pyrosequencing.
Understanding the molecular mechanisms involved in the development of porcine embryos prior to implantation is critical to the success of reproductive technologies such as embryo culture and transfer. Our current understanding of the gene expression profile for early development in the porcine embryo is limited. The purpose of this study was to use the Roche 454 deep sequencing platform to generate a massive EST dataset for the enrichment of previously unannotated genes expressed during the development of preimplantation porcine embryos. This information will eventually be used to establish a microarray platform for the study of differential gene expression at each specific developmental stage: GV, MII, 2-cell, 4-cell, 8-cell, morula and blastocyst under the activities of the EmbryoGENE NSERC Strategic Network.