不同保存方法对牛卵母细胞总RNA和cDNA品质的影响  

Effects of Different Methods on Preserving total RNA and cDNA of Bovine Oocytes

作　　者：易康乐[1] 李纯锦[2] 陈露[2] 孙燕玲[2] 燕海峰[1] 刘莹莹[1] 石德顺[3] 周虚[3] 

机构地区：[1]湖南省畜牧兽医研究所,湖南长沙410131 [2]吉林大学畜牧兽医学院,吉林长春130062 [3]广西大学动物繁殖研究所,广西南宁530005

出　　处：《家畜生态学报》2011年第3期18-21,共4页Journal of Domestic Animal Ecology

基　　金："863"计划项目(2008AA101003);国家科技支撑计划项目(2008BADB2B09-08)

摘　　要：实时荧光定量RT-PCR是检测低丰度mRNA的敏感方法,也是精确定量RNA拷贝数的有效方法,它的出现使得在少量细胞中定量基因的表达量,在小量基因组中分析基因突变和等位基因缺失成为可能。为了使这种检测技术更加稳定可靠,就必须在检测步骤的每一环节做到精确无误,而总mRNA的提取质量就显得十分重要。研究选择了牛MⅡ期的卵母细胞作为研究对象,对不同保存方法进行了检测和比较,结果显示,-196℃液氮和-80℃冰箱保存24 h的卵母细胞,总RNA品质并未明显下降;在-20℃或-80℃环境中长时间保存cDNA模板都会造成模板品质的下降。Real-time polymerase chain reaction（PCR） in combination with reverse transcription（RT） provides a powerful tool for accurate quantification of RNA copy numbers and has opened the way to the study of subtle modulations of gene expression in small numbers of cells,as well as small-scale genetic analyses aimed at establishing chromosome numbers,the presence of mutations,or allele dropout.The reliability of these measurements,however,depends on the accuracy of each step.Effects of different methods on preserving total RNA and cDNA of bovine oocytes（MⅡ） were studied.The results indicated that the quality of total RNA in-196℃ and-80℃ temperature were not significantly different than those in the control.However,the quality of cDNA in-20℃ or-80℃ temperature would be lost.

关 键 词：牛卵母细胞 总RNA CDNA 

分 类 号：S811.5[农业科学—畜牧学]