Separation of X- and Y-chromosome-bearing murine sperm by free-flow electrophoresis: evaluation of separation using PCR.
Zoolog Sci
; 9(3): 601-6, 1992 Jun.
Article
en En
| MEDLINE
| ID: mdl-1369423
The effectiveness of separation of murine X- and Y-bearing sperm by free-flow electrophoresis was evaluated by the polymerase chain reaction (PCR). The ratio of X- and Y-bearing sperm from cauda epididymis was analyzed before and after free-flow electrophoresis. A Y-chromosome-specific sequence (pY353/B) and an autosomal sequence (myogenin) were used to estimate the ratio between X- and Y-sperm in the separated fractions. Cauda epididymal mice sperm were separated into two peak fractions under the electric field. Each peak fraction contained sperm of normal shape, however, the motility of the sperm was extremely diminished after separation by electrophoresis. DNA was extracted from 10(4) sperm from each fraction and from the unseparated sperm, and Y-chromosome specific PCR was performed. The PCR experiment revealed that fraction No. 16 (the peak near the cathode) was a Y-sperm rich fraction, whereas fraction No. 22 (the peak near the anode) was a Y-sperm poor one. These results suggested that murine X- and Y-sperm could be successfully separated by free-flow electrophoresis. Analysis of the chromosome-specific sequence by PCR was demonstrated to be a direct and adequate method to evaluate the separation of X- and Y-sperm.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Espermatozoides
/
Cromosoma X
/
Cromosoma Y
/
Separación Celular
/
Reacción en Cadena de la Polimerasa
Límite:
Animals
Idioma:
En
Revista:
Zoolog Sci
Asunto de la revista:
BIOLOGIA
Año:
1992
Tipo del documento:
Article
País de afiliación:
Japón
Pais de publicación:
Japón