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Sexing of in vitro produced ovine embryos by duplex PCR

Mara, Laura and Pilichi, Susanna and Sanna, A. and Accardo, Carla and Chessa, Bernardo and Chessa, F. and Dattena, Maria and Bomboi, Giovanni Cristoforo and Cappai, Pietro (2004) Sexing of in vitro produced ovine embryos by duplex PCR. Molecular Reproduction and Development, Vol. 69 (1), p. 35-42. eISSN 1098-2795. Article.

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DOI: 10.1002/mrd.20147

Abstract

The aim of this article was to develop a fast and easy duplex polymerase chain reaction (PCR) method, for sex determination of ovine in vitro produced embryos prior to implantation. We tested the approach with 107 samples of autosomal cells (oviductal sheep cells and male lamb fibroblasts), divided into three groups for each sex according to the number of cells employed (30, 5, 2, respectively). We then used the test on 21 embryos at blastocyst stage. On the same day the embryos were transferred in pairs into 11 recipient synchronized ewes. The PCR utilized two different sets of primers: the first pair recognized a bovine Y-chromosome-specific sequence (SRY), that showed 100% homology with the corresponding sequence of the ovine Y-chromosome and is amplified in males only. The second pair recognized the bovine 1.715 satellite DNA (SAT) which was amplified in all ovine samples but, when submitted to the GenBank database did not show homology with any of the reported ovine sequences. However, after sequencing, ovine amplification product showed 98% homology with the bovine specific satellite sequence. The autosomal samples were amplified with 85.0% efficiency and 91.2% accuracy, while amplification was successful with all 21 embryos (100% efficiency). Eight lambs were born and the sex as determined by PCR corresponded to the anatomical sex in seven (87.5% accuracy). These results confirm that this method can be applied in ovine breeding programs to manipulate sex ratio of offspring.

Item Type:Article
ID Code:2509
Status:Published
Refereed:Yes
Uncontrolled Keywords:Sex determination, sheep, PCR
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/05 Malattie infettive degli animali domestici
Area 07 - Scienze agrarie e veterinarie > VET/02 Fisiologia veterinaria
Divisions:001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
002 Altri enti e centri di ricerca del Nord Sardegna > Istituto zootecnico e caseario per la Sardegna, Olmedo
001 Università di Sassari > 01 Dipartimenti > Biologia animale
Publisher:Wiley-Liss
eISSN:1098-2795
Deposited On:18 Aug 2009 10:07

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