Assessment of MDA efficiency for genotyping using cloned embryo biopsies
| Autor: | Paola Mariani, Irina Lagutina, Elena Genzini, Giovanna Lazzari, John L. Williams, Andrea Lauri, Cesare Galli, Francesco Braga |
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| Přispěvatelé: | Lauri A, Lazzari G, Galli C, Lagutina I, Genzini E, Braga F, Mariani P, Williams JL |
| Rok vydání: | 2013 |
| Předmět: |
Genotype
Genotyping Techniques Biopsy Cloning Organism Biology Embryo genotyping Genome Amplification quality assessment 03 medical and health sciences Multiple displacement amplification 0302 clinical medicine Embryo viability Genetics Animals Genotyping Alleles 030304 developmental biology Whole Genome Amplification 0303 health sciences 030219 obstetrics & reproductive medicine EMBRYO Embryo Sequence Analysis DNA Nucleic acid amplification technique Embryo Mammalian Molecular biology Whole genome amplification genotyping Cattle Nucleic Acid Amplification Techniques |
| Zdroj: | Genomics; Vol 101 |
| ISSN: | 0888-7543 |
| DOI: | 10.1016/j.ygeno.2012.09.002 |
| Popis: | The possibility to genotype embryos prior to implantation would have advantages for increasing the speed of selection of cattle. Reliable genotyping requires more DNA than can be obtained from biopsies of embryos, if they are to remain viable. Multiple displacement amplification (MDA) is a whole genome amplification technique used to increase the amount of DNA from biopsies for analysis. Reduced genome coverage resulting in Allele Drop Out (ADO) at heterozygous loci or missing genotypes are drawbacks of MDA.The present article describes the correlation between the input DNA quantity or embryo biopsy size and MDA success. Missing genotypes and ADO drastically increased when fewer than 30–40 cells or the genomic equivalents were used. However, embryo viability was found to be reduced if biopsied with more than 10 cells. Therefore, in vitro cell culture was investigated as a means to increase the number of cells available and the genotyping reliability. |
| Databáze: | OpenAIRE |
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