COMET ASSAY AS DNA INTEGRITY TEST FOR BOAR SPERMATOZA FOLLOWING PRESERVATION WITH BTS DILUENT AT 18° CB.C. DAS, S SANYAL, K.M. BO A RBARUA1-11, P.K. DAS, P.R. GHOSH, S. NASKAR1, M.H. KHAN, A.B. DAS, S. BANDOPADHYAV
The assessment of DNA integrity influenced by sperm ageing, which is manifested in DNA double strand break was performed under neutral condition using cornet assay. In the present investigation an attempt has been made to use modified neutral comet assay which also could be termed as single cell gel electrophoresis to assess DNA integrity of boar spermatozoa during liquid storage for 24, 48 and 72 hours at 18Â° C. In the protocol 2% 133-mercaptoethanol has been used prior to the lysis procedure for removing nuclear protein. A total of six boars, three each from Hampshire and Crossbred (Hampshire X Khasi local) taken for this study. Ejaculates sample taken for cornet at 0 hour and then preserved for 24, 48 and 72 hours at 18Â° C. At 0 hour comet percentage was minimum and as the time advances it increases along with time. There was no individual variation among the boars and between the breeds. Study indicates there was elevated level of DNA damage at prolong storage (72 hours) and it seems that long term storage can affect genomic integrity of boar spermatozoa. Therefore, screening of sperm DNA damage may be used as an additional test of sperm function that can have diagnostic value in practice.
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