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This has the potential to considerably improve cattle breeding, genetic resource management and particularly the efficiency of dairy and meat production. However, the broad use of sexed semen will depend on availability, price, fertilizability and in particular the actual sorting purity of sperm doses. To validate the accuracy of sperm sexing in <jats:italic>Bos taurus<\/jats:italic>, we have developed a simple, fast and reliable dual colour fluorescence <jats:italic>in\u00a0situ<\/jats:italic> hybridization (FISH) test, where Y\u2010bearing spermatozoa are identified by a DNA fragment hybridizing to a large pericentromeric repetitive DNA block on the bovine Y chromosome (locus <jats:italic>DYZ1<\/jats:italic>, Yp13\u2013q12). To avoid an underestimation of Y signals, we used a second DNA probe identifying a large subcentromeric block of complex repetitive DNA on the bovine autosome 6 (locus <jats:italic>D6Z1<\/jats:italic>, 6q12\u201315) as a positive control. Bovine sperm were fixed with methanol:acetic acid and denatured by simply immersing in 3\u2003<jats:sc>m<\/jats:sc> NaOH, yielding consistent hybridization results and good preservation of sperm morphology. The FISH protocol was evaluated on unsorted sperm as well as on sperm samples sexed using the Beltsville technology, which separates X\u2010 and Y\u2010bearing spermatozoa by staining with Hoechst 33342 and flow sorting according to their DNA content (<jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"#b12\">Johnson <jats:italic>et\u00a0al.<\/jats:italic> 1987<\/jats:ext-link>).<\/jats:p>","DOI":"10.1111\/j.1439-0388.2005.00488.x","type":"journal-article","created":{"date-parts":[[2005,2,24]],"date-time":"2005-02-24T16:19:55Z","timestamp":1109261995000},"page":"22-27","source":"Crossref","is-referenced-by-count":20,"title":["Validation of sperm sexing in the cattle (<i>Bos taurus<\/i>) by dual colour fluorescence <i>in\u00a0situ<\/i> hybridization"],"prefix":"10.1111","volume":"122","author":[{"given":"F.A.","family":"Habermann","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Winter","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"I.","family":"Olsaker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P.","family":"Reichert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"Fries","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,2,24]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/25.17.3389"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0093-691X(99)00219-8"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0442.1994.tb00113.x"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1159\/000134361"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.20016"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00218904"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(199908)53:4<407::AID-MRD6>3.0.CO;2-O"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0320(20010201)43:2<101::AID-CYTO1024>3.0.CO;2-8"},{"key":"e_1_2_9_10_1","first-page":"283","volume-title":"Cytogenet Cell Genet","author":"ISCNDB 2000","year":"2001"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0388.2004.00455.x"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0093-691X(99)00220-4"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.1120170303"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2795(199812)51:4<390::AID-MRD5>3.0.CO;2-F"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0093-691X(99)00193-4"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0888-7543(90)90478-D"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1186\/1297-9686-33-1-89"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1530\/rep.0.1210541"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1071\/RD02026"},{"key":"e_1_2_9_20_1","first-page":"365","article-title":"Primer3 on the www for general users and for biologist programmers","volume":"132","author":"Rozen S.","year":"2000","journal-title":"Methods Mol. 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