title: Transcriptome Profiling of In-Vivo Produced Bovine Pre-implantation Embryos Using Two-color Microarray Platform.
abstract: Early embryonic loss is a large contributor to infertility in cattle. Moreover, bovine becomes an interesting model to study human preimplantation embryo development due to their similar developmental process. Although genetic factors are known to affect early embryonic development, the discovery of such factors has been a serious challenge. Microarray technology allows quantitative measurement and gene expression profiling of transcript levels on a genome-wide basis. One of the main decisions that have to be made when planning a microarray experiment is whether to use a one- or two-color approach. Two-color design increases technical replication, minimizes variability, improves sensitivity and accuracy as well as allows having loop designs, defining the common reference samples. Although microarray is a powerful biological tool, there are potential pitfalls that can attenuate its power. Hence, in this technical paper we demonstrate an optimized protocol for RNA extraction, amplification, labeling, hybridization of the labeled amplified RNA to the array, array scanning and data analysis using the two-color analysis strategy.
authors.affiliation: Department of Agricultural, Food and Nutritional Science, University of Alberta.
authors.forename: Reza
authors.initials: R
authors.lastname: Salehi
authors.affiliation: Department of Agricultural, Food and Nutritional Science, University of Alberta.
authors.forename: Stephen C M
authors.initials: SC
authors.lastname: Tsoi
authors.affiliation: Livestock Research Branch, Alberta Agriculture and Forestry.
authors.forename: Marcos G
authors.initials: MG
authors.lastname: Colazo
authors.affiliation: Department of Agricultural, Food and Nutritional Science, University of Alberta; Livestock Research Branch, Alberta Agriculture and Forestry.
authors.forename: Divakar J
authors.initials: DJ
authors.lastname: Ambrose
authors.affiliation: Laboratory of Functional Genomics of Early Embryonic Development, Université Laval.
authors.forename: Claude
authors.initials: C
authors.lastname: Robert
authors.affiliation: Department of Agricultural, Food and Nutritional Science, University of Alberta; mkdyck@ualberta.ca.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Dyck
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D047108:Embryonic Development; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D020869:Gene Expression Profiling; D008855:Microscopy, Electron, Scanning; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011247:Pregnancy; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012984:Software
chemical_list: D005456:Fluorescent Dyes; D012313:RNA