Impact of whole-genome amplification on the reliability of pre-transfer cattle embryo breeding value estimates
| Autor: | Habib A. Shojaei Saadi, Dominic Gagné, Christian Vigneault, Mehdi Sargolzaei, Claude Robert, Eric Fournier, Patrick Blondin, Béatrice de Montera, J. P. Chesnais |
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| Rok vydání: | 2014 |
| Předmět: |
Genotyping
Genotyping Techniques Population Loop-mediated isothermal amplification Biology Breeding Genomic breeding value Polymorphism Single Nucleotide Bovine early embryo 03 medical and health sciences 0302 clinical medicine Multiple displacement amplification Genetics Pre-Implantation genetic diagnosis Animals Whole-genome amplification education Cells Cultured 030304 developmental biology Whole Genome Amplification 0303 health sciences education.field_of_study 030219 obstetrics & reproductive medicine Genome Genotype imputation Methodology Article Reproducibility of Results DNA Genomics Fibroblasts Embryo Mammalian genomic DNA Cattle Female Embryo biopsy Imputation (genetics) Biotechnology |
| Zdroj: | BMC Genomics |
| ISSN: | 1471-2164 |
| Popis: | Background Genome-wide profiling of single-nucleotide polymorphisms is receiving increasing attention as a method of pre-implantation genetic diagnosis in humans and of commercial genotyping of pre-transfer embryos in cattle. However, the very small quantity of genomic DNA in biopsy material from early embryos poses daunting technical challenges. A reliable whole-genome amplification (WGA) procedure would greatly facilitate the procedure. Results Several PCR-based and non-PCR based WGA technologies, namely multiple displacement amplification, quasi-random primed library synthesis followed by PCR, ligation-mediated PCR, and single-primer isothermal amplification were tested in combination with different DNA extractions protocols for various quantities of genomic DNA inputs. The efficiency of each method was evaluated by comparing the genotypes obtained from 15 cultured cells (representative of an embryonic biopsy) to unamplified reference gDNA. The gDNA input, gDNA extraction method and amplification technology were all found to be critical for successful genome-wide genotyping. The selected WGA platform was then tested on embryo biopsies (n = 226), comparing their results to that of biopsies collected after birth. Although WGA inevitably leads to a random loss of information and to the introduction of erroneous genotypes, following genomic imputation the resulting genetic index of both sources of DNA were highly correlated (r = 0.99, P |
| Databáze: | OpenAIRE |
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