摘要
目的 :构建AFPmRNA的realtimeRT PCR标准定量模板。方法 :提取肝瘤细胞株BEL740 2细胞总RNA ,进行RT PCR扩增并纯化AFP基因片段 ,与PMD 1 8T载体连接构建重组质粒。结果 :重组质粒的阳性克隆效率为81 .8% ,经酶切鉴定 ,目的基因片段已插入PMD 1 8T载体内。结论 :成功构建了AFPmRNA的标准定量模板 ,可应用于该基因的realtimeRT
Aim:To construct standard quantitative templates of AFP mRNA for real time RT PCR.Methods:Total mRNA of BEL7402 cells was extracted. The RT PCR products of AFP mRNA were purified and combined with PMD 18T vectors.Results:The positive efficiency of recombinant plasmids were 81.8%. It was proved that the aimed gene had been cloned into PMD 18T vectors by enzyme digest.Conclusion:The standard quantitative templates of AFP mRNA were successfully constructed and could be used in real time RT PCR for detecting the expression levels of AFP mRNA.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2003年第2期228-229,共2页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省科技攻关基金资助项目0 2 2 4 6 30 0 32
