南方农业学报2011，Vol.42Issue(8)：984-986,3.
多重PCR鉴定水牛胚胎性别的研究
Sex identification of buffalo embryo using multiple-PCR
摘要
Abstract
[Objective]The present experiment was conducted to develop a fast and accurate PCR based method for sex identification of buffalo embryos by PCR for providing the basis of optimizing multiple nested PCR method. [Method ]Three methods to extract DNA from buffalo blastomere, including thermal lyses, proteinase K+DTT and DTT+Tween-20, were used and the isolated DNAs were comparatively analyzed for their amplification efficiency using primers based on G3PDH gene of buffalo. The embryonic DNA was amplified by using pairs of detection primer and an internal primer in multiple-PCR amplification. [ Result ]The results indicated that DTT+Tween-20 methods were the best for cracking embryonic DNAs and gave 97.1% detectable rate, and 100% embryo identification rate in sex identification using multiple-PCR amplification. [ Conclusion ]The embryo sample must be initially treated with DTT+Tween-20 for the lyses of cells so as to completely extract the DNA followed by amplification using multiple-PCR method. The present method has simplified operating steps and therefore, many samples can be processed in less time compared to the other sex identification methods.关键词
水牛胚胎/性别鉴定/多重PCRKey words
buffalo embryo/ sex identification/ multiple-PCR分类
农业科技引用本文复制引用
付强,郑海英,陈焕华,张明,卢克焕..多重PCR鉴定水牛胚胎性别的研究[J].南方农业学报,2011,42(8):984-986,3.基金项目
广西青年科学基金项目(2011GXNSFB018028) （2011GXNSFB018028）
广西自然科学基金项目(桂科自0991047) （桂科自0991047）
