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The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> and, consequently, its very small amounts in maternal bloodstream. Circulating cell\u2010free foetal <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> (ccff<jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content>) encompasses short nucleotide fragments (300\u2013600\u00a0bp) in maternal circulation. The aim of this study was to assess this non\u2010invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non\u2010pregnant heifer was the controls. Extraction of foetal <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> was accomplished by three different methods: using tubes with silicone membranes, a single\u2010tube extraction without silicone membranes and phenol\u2013chloroform extraction. Following each extraction method, foetal <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> was amplified using <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> and real\u2010time <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> with both <jats:styled-content style=\"fixed-case\">bAML<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">TSPY<\/jats:styled-content> primers in a separate reaction. Positive results were obtained only after amplification of foetal <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> extracted with a single\u2010tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> test was 90% in Group I, but the technique failed to detect male foetuses in Group <jats:styled-content style=\"fixed-case\">II<\/jats:styled-content>. The real\u2010time <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> test sensitivity in Group I was 90% and in Group <jats:styled-content style=\"fixed-case\">II<\/jats:styled-content> 91.6%.<\/jats:p>","DOI":"10.1111\/rda.13193","type":"journal-article","created":{"date-parts":[[2018,5,9]],"date-time":"2018-05-09T04:50:02Z","timestamp":1525841402000},"page":"947-954","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Bovine foetal sex determination\u2014Different <scp>DNA<\/scp> extraction and amplification approaches for efficient livestock production"],"prefix":"10.1111","volume":"53","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6084-7321","authenticated-orcid":false,"given":"M","family":"Ristanic","sequence":"first","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lj","family":"Stanisic","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M","family":"Maletic","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"U","family":"Glavinic","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"V","family":"Draskovic","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N","family":"Aleksic","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Z","family":"Stanimirovic","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine University of Belgrade  Belgrade Serbia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2018,5,8]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/pd.69"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10038-003-0084-5"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(03)12191-5"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2008.112797"},{"key":"e_1_2_9_6_1","first-page":"4455","article-title":"Sexing bovine pre\u2010implantation embryos using the polymerase chain reaction: A model for human embryo sexing","volume":"11","author":"Cenariu M.","year":"2014","journal-title":"African Journal of Biotechnology"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2003.024893"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(199911)54:3<209::AID-MRD1>3.0.CO;2-6"},{"key":"e_1_2_9_9_1","doi-asserted-by":"crossref","first-page":"1607","DOI":"10.1093\/clinchem\/47.9.1607","article-title":"Effects of blood\u2010processing protocols on fetal and total DNA quantification in maternal plasma","volume":"47","author":"Chiu R. 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