Short Communication

Microsatellite analysis of cryopreserved stallion semen stored on FTA(R) paper : research communication

M.L. Schulman, C.K. Harper, E. Bell, A. Nel, A.J. Guthrie
Journal of the South African Veterinary Association | Vol 73, No 4 | a592 | DOI: https://doi.org/10.4102/jsava.v73i4.592 | © 2002 M.L. Schulman, C.K. Harper, E. Bell, A. Nel, A.J. Guthrie | This work is licensed under CC Attribution 4.0
Submitted: 06 July 2002 | Published: 06 July 2002

About the author(s)

M.L. Schulman,
C.K. Harper,
E. Bell,
A. Nel,
A.J. Guthrie,

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Abstract

The aim of this study was to establish and validate a method to permit microsatellite analysis of DNA profiles obtained from frozen-thawed stallion sperm cells. This would provide reliable and accurate verification of the identification of a semen donor. Ejaculates from 5 pony stallions were collected, processed and frozen in 0.5 m plastic straws. Aliquots of 100 m of the frozen-thawed semen thus obtained were either placed directly, or diluted (1 : 10 ; 1 : 100 ; and 1 : 1000) and placed on slides of FTA(R) paper. Similarly, blood samples obtained from each of the stallions were placed onto slides of FTA(R) paper. A punch was removed from each sample after drying. Each sample was mixed with FTA(R) purification reagent, Dithiothreitol and Proteinase K before incubation and processing. All samples were processed with a set of 13 microsatellite markers. Further analysis permitted a comparison of the DNA profiles of the frozen-thawed semen and the blood samples. A full profile of markers was obtained from the 1 : 10 and 1 : 100 dilutions of the frozen-thawed semen samples as well as from the blood samples. The DNA profiles from the frozen-thawed semen and blood samples obtained from the stallions matched in all cases.

Keywords

Cryopreserved Semen; DNA Microsatellites; Stallion

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