Detection of Y‐bearing porcine spermatozoa by in situ hybridization using digoxigenin‐labeled, porcine male‐specific DNA probe produced by polymerase chain reaction

摘要: This study was carried out to determine whether Y-bearing porcine spermatozoa could be detected by in situ hybridization using a digoxigenin (Dig)-labelled DNA probe specific the Y chromosome produced polymerase chain reaction (PCR). A conventional PCR (with Dig-dUTP) performed set of oligonucleotide primers (5′-AAGTGGTCAGCGTGTCCATA-3′ and 5′-TTTCTCCTGTATCCTCCTGC-3′) for 236 bp fragment male-specific sequence 1.25 × 104 template white blood cells obtained from boar. When fluorescence with Dig-labelled applied metaphase spreads prepared both boar gilts, fluorescein signal only on long arm chromosome. In addition, immunocytochemical detection alkaline phosphatase-labeled anti-Dig sperm nuclei pretreated dithiothreitol cells; 51% 96% were labelled, whereas none gilts labelled probe. The results indicated that made possible direct visualization hybridization. © 1995 Wiley-Liss, Inc.