Quantification of Y chromosome bearing spermatozoa of cattle using in situ hybridization.
An easy assay for quantification of Y chromosome-bearing sperm (Y-sperm) is needed, especially to monitor sperm separation techniques. In the present study a tritiated bovine male-specific DNA fragment was tested for identification of Y-sperm by in situ hybridization. A protocol for in situ hybridization to bovine sperm was developed and used to study the proportion of Y-sperm of 12 bulls. The usefulness of the method in optimization of sperm separation procedures is illustrated through analysis of fractions of sperm separated by Percoll density gradient centrifugation.
Schwerin
M
M
Department of Molecular Genetics, Research Center of Animal Production Dummerstorf-Rostock, Federal Republic of Germany.
Blottner
S
S
Thomsen
P D
PD
Roschlau
D
D
Brockmann
G
G
D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000235:genetics*; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D004247:DNA / Q000302:isolation & purification*; D015342:DNA Probes; D004277:DNA, Single-Stranded; D008297:Male; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012727:Sex Characteristics; D013094:Spermatozoa; D014018:Tissue Distribution; D014998:Y Chromosome
D015342:DNA Probes; D004277:DNA, Single-Stranded; D004247:DNA