Verification of caprine sexed-separated spermatozoa by Real Time PCR

Somarny W.W.M.Z., . and Aslinda K., . and Md. Tasol S., . and Mohd. Hifzan R., . and Nasir J.M., . and Mohd Hafiz A.W., . and Ab Rashid B., . and Musaddin K., . Verification of caprine sexed-separated spermatozoa by Real Time PCR. pp. 73-83. ISSN 1394-3227

Sexed spermatozoa could contribute to increasing profitability of the dairy and beef industries worldwide by producing offspring of predetermined sex. Various methods have been designed to separate sperm population into X- and Y-chromosome bearing spermatozoa. In this study the caprine semen sample from Savanna (n2) and Boer (n4) breeds were sex-separated using free flow electrophoresis. In order to determine the efficiency of sex-separation procedure a rapid and accurate technique real time PCR (qPCR) was used to validate the X- and Y-chromosome bearing spermatozoa following separation. The primers and probes were developed to detect the X- and Y- chromosome bearing spermatozoa of Capra hircus based on ZFX and SRY genes respectively using TaqMan qPCR assay. The amplification of ZFX and SRY genes produced a single fragment of 93 bp and 137 bp respectively. The primers used in separated spermatozoa by free-flow electrophoresis technique showed that the mean ratio of X- and Y-chromosomes bearing spermatozoa from the cathodic and anodic fractions were 36:64 and 44:56 respectively compared to unseparated semen was 51:49. Thus these primers can be used to validate the gene copy of X- and Y-chromosome bearing spermatozoa in caprine semen.

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Abstract

Sexed spermatozoa could contribute to increasing profitability of the dairy and beef industries worldwide by producing offspring of predetermined sex. Various methods have been designed to separate sperm population into X- and Y-chromosome bearing spermatozoa. In this study the caprine semen sample from Savanna (n2) and Boer (n4) breeds were sex-separated using free flow electrophoresis. In order to determine the efficiency of sex-separation procedure a rapid and accurate technique real time PCR (qPCR) was used to validate the X- and Y-chromosome bearing spermatozoa following separation. The primers and probes were developed to detect the X- and Y- chromosome bearing spermatozoa of Capra hircus based on ZFX and SRY genes respectively using TaqMan qPCR assay. The amplification of ZFX and SRY genes produced a single fragment of 93 bp and 137 bp respectively. The primers used in separated spermatozoa by free-flow electrophoresis technique showed that the mean ratio of X- and Y-chromosomes bearing spermatozoa from the cathodic and anodic fractions were 36:64 and 44:56 respectively compared to unseparated semen was 51:49. Thus these primers can be used to validate the gene copy of X- and Y-chromosome bearing spermatozoa in caprine semen.

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Item Type:	Article
AGROVOC Term:	Spermatozoa
AGROVOC Term:	Electrophoresis
AGROVOC Term:	PCR
AGROVOC Term:	Caprinae
AGROVOC Term:	Goats
AGROVOC Term:	Cattle
AGROVOC Term:	Dairy industry
AGROVOC Term:	beef
AGROVOC Term:	Profitability
Geographical Term:	Malaysia
Depositing User:	Ms. Suzila Mohamad Kasim
Last Modified:	26 Apr 2025 14:37
URI:	http://webagris.upm.edu.my/id/eprint/21518

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