Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine
The aim of the present study was to verify the presence of X- and Y-chromosome spermatozoa after separation with swimming speed using oestrus cows vagina mucus, Percoll discontinuous gradient (45 to 90%) and swim-up using TALP medium. The nested polymerase chain reaction (PCR) was used to determine...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Academic Journals
2011
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| Online Access: | http://psasir.upm.edu.my/id/eprint/25345/1/25345.pdf |
| _version_ | 1796970740217544704 |
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| author | Abdul Malik, Haron, Abd Wahid Yusoff, Rosnina Bukar, Muhammad Modu Kasim, Azhar Mohd Yusoff, Md Sabri |
| author_facet | Abdul Malik, Haron, Abd Wahid Yusoff, Rosnina Bukar, Muhammad Modu Kasim, Azhar Mohd Yusoff, Md Sabri |
| author_sort | Abdul Malik, |
| collection | UPM |
| description | The aim of the present study was to verify the presence of X- and Y-chromosome spermatozoa after separation with swimming speed using oestrus cows vagina mucus, Percoll discontinuous gradient (45 to 90%) and swim-up using TALP medium. The nested polymerase chain reaction (PCR) was used to determine X- and Y-chromosome bearing spermatozoa after separation. The primers for PCR were designed using amelogenin cDNA sequence with 329 and 266 bp for X- and Y-bearing chromosome spermatozoa, respectively. The motility was analyzed using computer assisted sperm analyser, whereas the membrane integrity was analyzed using hypo-osmotic swelling test (HOST). Results were confirm by the absence of single band, either for X- or Y-chromosome. Inversely, the double band indicating that the spermatozoa cannot be separated was observed. The percentage of X-chromosome bearing spermatozoa in the swimming speed using oestrus cows vagina mucus media, Percoll discontinuous gradient and swim up methods were 58.33, 44.33 and 50%, respectively. Statistically, both percentages were significantly different (P<0.001) as compared to the theoretical ratio (50:50). Spermatozoa motility, membrane integrity and concentration before and after separation were also significantly different (P<0.05). This study shows that although swimming speed using oestrus cow’s vagina mucus media may be used to separate X- and Y-chromosome bearing spermatozoa in bulls, the results however, require further investigation. |
| first_indexed | 2024-03-06T08:02:33Z |
| format | Article |
| id | upm.eprints-25345 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T08:02:33Z |
| publishDate | 2011 |
| publisher | Academic Journals |
| record_format | dspace |
| spelling | upm.eprints-253452016-04-27T03:56:07Z http://psasir.upm.edu.my/id/eprint/25345/ Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine Abdul Malik, Haron, Abd Wahid Yusoff, Rosnina Bukar, Muhammad Modu Kasim, Azhar Mohd Yusoff, Md Sabri The aim of the present study was to verify the presence of X- and Y-chromosome spermatozoa after separation with swimming speed using oestrus cows vagina mucus, Percoll discontinuous gradient (45 to 90%) and swim-up using TALP medium. The nested polymerase chain reaction (PCR) was used to determine X- and Y-chromosome bearing spermatozoa after separation. The primers for PCR were designed using amelogenin cDNA sequence with 329 and 266 bp for X- and Y-bearing chromosome spermatozoa, respectively. The motility was analyzed using computer assisted sperm analyser, whereas the membrane integrity was analyzed using hypo-osmotic swelling test (HOST). Results were confirm by the absence of single band, either for X- or Y-chromosome. Inversely, the double band indicating that the spermatozoa cannot be separated was observed. The percentage of X-chromosome bearing spermatozoa in the swimming speed using oestrus cows vagina mucus media, Percoll discontinuous gradient and swim up methods were 58.33, 44.33 and 50%, respectively. Statistically, both percentages were significantly different (P<0.001) as compared to the theoretical ratio (50:50). Spermatozoa motility, membrane integrity and concentration before and after separation were also significantly different (P<0.05). This study shows that although swimming speed using oestrus cow’s vagina mucus media may be used to separate X- and Y-chromosome bearing spermatozoa in bulls, the results however, require further investigation. Academic Journals 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/25345/1/25345.pdf Abdul Malik, and Haron, Abd Wahid and Yusoff, Rosnina and Bukar, Muhammad Modu and Kasim, Azhar and Mohd Yusoff, Md Sabri (2011) Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine. African Journal of Biotechnology, 10 (85). art. no. F158CC438333. pp. 19796-19801. ISSN 1684–5315 http://www.academicjournals.org/journal/AJB/article-abstract/F158CC438333 |
| spellingShingle | Abdul Malik, Haron, Abd Wahid Yusoff, Rosnina Bukar, Muhammad Modu Kasim, Azhar Mohd Yusoff, Md Sabri Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine |
| title | Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine |
| title_full | Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine |
| title_fullStr | Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine |
| title_full_unstemmed | Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine |
| title_short | Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine |
| title_sort | verification of x and y spermatozoa separation by nested polymerase chain reaction pcr motility and membrane integrity in bovine |
| url | http://psasir.upm.edu.my/id/eprint/25345/1/25345.pdf |
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