Rapid and simultaneous detection of chromosome Y- and 1-bearing porcine spermatozoa by fluorescence in situ hybridization
作者： Tatsuo Kawarasaki , Masaru Sone , Mitsutoshi Yoshida , Kimio Bamba
DOI: 10.1002/(SICI)1098-2795(199604)43:4<548::AID-MRD18>3.0.CO;2-V
关键词: Biology 、 Chromosome 、 Y chromosome 、 Hybridization probe 、 Sperm 、 Molecular biology 、 Digoxigenin 、 Fluorescence in situ hybridization 、 Polymerase chain reaction 、 DNA 、 Developmental biology 、 Genetics 、 Cell biology
摘要: This study was carried out to develop a rapid and simultaneous detection system of chromosome Y- 1-bearing porcine spermatozoa by fluorescence in situ hybridization (FISH). Chromosome 1-specific DNA probes were produced polymerase chain reaction with digoxigenin (Dig)- or biotin-dUTP. The probe mixture labeled Y-chromosome applied the preparation, immediately denatured at 75°C for 8 min, hybridized 5 min 37°C overall FISH steps done within few hours. When double Dig-labeled Y-specific biotin-labeled sperm nuclei pretreated dithiothreitol, average 50.9% had Dig-signal, 99.2% biotin-signal 0.3% showed no signal. putative rate Y-bearing ranged from 49.8% 52.8% among boars 51.0%. results indicated that PCR made possible more accurate assessment spermatozoa. © 1996 Wiley-Liss, Inc.
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