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Assessment of MDA efficiency for genotyping using cloned embryo biopsies
- Source :
- Genomics; Vol 101
- Publication Year :
- 2013
- Publisher :
- Elsevier BV, 2013.
-
Abstract
- 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.
- Subjects :
- 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
Subjects
Details
- ISSN :
- 08887543
- Volume :
- 101
- Database :
- OpenAIRE
- Journal :
- Genomics
- Accession number :
- edsair.doi.dedup.....2fbc20d59f8085586135c1f4747b1011