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KMID : 1005220100250020111
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Journal of Embryo Transfer
2010 Volume.25 No. 2 p.111 ~ p.116
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Evaluation of DNA Fragments on Boar Sperm by Ligation-mediated Quantitative Real Time PCR
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Lee Eun-Soo
Choi Sun-Gyu
Yang Jae-Hun
Bae Mun-Sook
Park Jin-Young
Park Hong-Min
Han Tae-Kyu
Hwang You-Jin
Kim Dae-Young
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Abstract
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Sperm chromatin integrity is essential for successful fertilization and development of an embryo. Reported here is a quantification of DNA fragments which is intimately associated with reproductive potential to provide one of criteria for sperm chromatin integrity. Three sperm populations were considered: CONTROL (no treatment), UV irradiation (48mW/, 1h) and (oxidative stress induced by hydrogen peroxide, 10 mM, 50 mM and 100 mM). DNA fragments in boar sperm were evaluated by using ligation-mediated quantitative real-time polymerase chain reaction (LM-qPCR) assay, which relies on real-time qPCR to provide a measure of blunt 5’ phosphorylated double strand breaks in genomic DNA. The results in agarose gel electrophoresis showed no significant DNA fragmentation and no dose-dependent response to . However, the remarkable difference in shape and position was observed in melting curve of LM-qPCR. This result supported that the melting curve analysis of LM-qPCR presented here, could be more sensitive and accurate than previous DNA fragmentation assay method.
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KEYWORD
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DNA fragmentation, ligation-mediated quantitative real-time polymerase chain reaction (LM-qPCR), boar sperm, sperm quality
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