Evaluation of DNA Fragments on Boar Sperm by Ligation-mediated Quantitative Real Time PCR
(주)코리아스칼라
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- 2016.04.01
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서지정보
ㆍ발행기관 : 한국수정란이식학회
ㆍ수록지정보 : Journal of Embryo Transfer / 25권 / 2호
ㆍ저자명 : 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
영어 초록
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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