猪精子RNA提取及cDNA质量检测  

The RNA extraction and detection of sperm cDNA quality in pig

作　　者：高波[1] 宋成义[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《黑龙江畜牧兽医》2014年第4期61-63,209,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31272406;31200920)

摘　　要：为了有效分离猪精子RNA,试验通过精子上游法获得纯净精子样本,采用改良TRIzol法提取精子总RNA,经核酸测定仪及琼脂糖凝胶电泳检测RNA的纯度和质量,RT-PCR检测相应体细胞标记基因E-Cadherin、CD4和c-kit。结果表明:研究建立的精子RNA提取法可获得高质量的RNA,浓度约为203 ng/μL,OD260/280值为1.99,并且无其他体细胞标记基因扩增产物,可用于猪精子mRNA的研究。To effectively isolate the sperm RNA in pig, the swim - up protocol was used to obtain the purified sperms. The total RNA from sperm was extracted using an improved Trizol method, and then the purity and quantity of RNA were determined using a nucleic acid tester and agarase gel electrophoresis. The RT - PCR was used to detect the E - Cadherin gene, CD4 gene and c - kit gene used as the marker genes correspond- ing to the somatic ceils. The results showed that the established extraction method for sperm RNA could obtain high - quality RNA, its concen- tration was approximately 203 ng/μL, and the OD260/250 ratio was 1.99. There were no other amplification products from the other marker genes of the somatic ceils. The results indicate that the extraction method can be used to study sperm mRNA in pig.

关 键 词：精子RNA CDNA 猪 

分 类 号：S828.34[农业科学—畜牧学] Q522[农业科学—畜牧兽医]