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Multiple genotype analysis and sexing of IVF bovine embryos
Year:
1996
Source of publication :
Theriogenology
Authors :
Dekel, Itzhak
;
.
Feldmesser, Ester
;
.
Hochman, Daniel
;
.
Ron, Micha
;
.
Shani, Moshe
;
.
Zaron, Yoel
;
.
Volume :
46
Co-Authors:
Hochman, D., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Zaron, Y., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Dekel, L., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Feldmesser, E., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Medrano, J.F., Department of Animal Science, University of California, Davis, CA, United States
Shani, M., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Ron, M., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Facilitators :
From page:
1063
To page:
1075
(
Total pages:
13
)
Abstract:
Twenty-one in vitro-fertilized bovine blastocysts were quartered, lysed and subjected to primer elongation preamplification (PEP) procedure, allowing for the analysis of up to 40 genotypes per quarter embryo. The quarter- embryos were sexed by polymerase chain reaction (PCR) using BRY.1, Bov97M and ZFX/ZFY loci, and then genotyped for k-casein, bovine leukocyte adhesion deficiency (BLAD) and microsatellite D9S1. The mitochondrial cytochrome B locus was used as an internal control with a 95% success rate. The PEP procedure amplified genomic fragments in 93% of all cases. The embryos were identified to be 11 males and 10 females. Sexing accuracy was 87% for BRY.1, 97% for ZFX/ZFY and 100% for Bov97M. False genotyping was due mostly to amplification of BRY.1 in the female embryos and to the nonamplification of the ZFY locus in the male embryos. The results indicate that the combined use of Bov97M and ZFX/ZFY loci is a highly accurate procedure for sexing bovine embryos. Genotyping for κ-casein, D9S1 and BLAD was successful in 94, 99 and 91% of assays, respectively. Sex ratios and allele frequencies of embryos for κ-casein, BLAD and D9S1 were all close to the observed frequencies in the Israeli Holstein population. These results support the conclusion that the genotyping of embryos is as accurate as that of mature animals. Thus, marker- assisted selection can be efficiently applied at the preimplantation embryo level for loci of economic importance.
Note:
Related Files :
animal cell
animal tissue
cattle
Female
Fertilization
gene amplification
genetic analysis
in vitro fertilization
sex ratio
Show More
Related Content
More details
DOI :
10.1016/S0093-691X(96)00271-3
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
21458
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:44
You may also be interested in
Scientific Publication
Multiple genotype analysis and sexing of IVF bovine embryos
46
Hochman, D., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Zaron, Y., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Dekel, L., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Feldmesser, E., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Medrano, J.F., Department of Animal Science, University of California, Davis, CA, United States
Shani, M., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Ron, M., Institute of Animal Science, Volcani Center, P.O.B 6, Bet-Dagan, Israel
Multiple genotype analysis and sexing of IVF bovine embryos
Twenty-one in vitro-fertilized bovine blastocysts were quartered, lysed and subjected to primer elongation preamplification (PEP) procedure, allowing for the analysis of up to 40 genotypes per quarter embryo. The quarter- embryos were sexed by polymerase chain reaction (PCR) using BRY.1, Bov97M and ZFX/ZFY loci, and then genotyped for k-casein, bovine leukocyte adhesion deficiency (BLAD) and microsatellite D9S1. The mitochondrial cytochrome B locus was used as an internal control with a 95% success rate. The PEP procedure amplified genomic fragments in 93% of all cases. The embryos were identified to be 11 males and 10 females. Sexing accuracy was 87% for BRY.1, 97% for ZFX/ZFY and 100% for Bov97M. False genotyping was due mostly to amplification of BRY.1 in the female embryos and to the nonamplification of the ZFY locus in the male embryos. The results indicate that the combined use of Bov97M and ZFX/ZFY loci is a highly accurate procedure for sexing bovine embryos. Genotyping for κ-casein, D9S1 and BLAD was successful in 94, 99 and 91% of assays, respectively. Sex ratios and allele frequencies of embryos for κ-casein, BLAD and D9S1 were all close to the observed frequencies in the Israeli Holstein population. These results support the conclusion that the genotyping of embryos is as accurate as that of mature animals. Thus, marker- assisted selection can be efficiently applied at the preimplantation embryo level for loci of economic importance.
Scientific Publication
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