脂肪组织RNA提取方法的改进  

Improvement of Methods for Extracting Total RNA from Adipose

作　　者：吴江维[1] 杨公社[1] 孙超[1] 

机构地区：[1]西北农林科技大学动物脂肪沉积与肌肉发育实验室,杨凌712100

出　　处：《生物技术通报》2005年第5期75-77,81,共4页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30471267);中国博士后科学基金项目

摘　　要：本试验以猪脂肪组织为材料,应用TRIzol、V-gene、H.Q.&.Q三种总RNA提取试剂盒分别提取脂肪组织RNA,发现直接按照试剂盒说明书操作提取的RNA效果不理想。通过反复试验,分别改进了提取过程中的部分操作步骤,结果表明:三种试剂盒操作方法改进后与改进前相比,提取RNA的纯度和得率表现为差异极显著(P<0.01);三种试剂盒操作方法改进后提取RNA的纯度差异不显著(P>0.05),但在得率方面,Trizol比其它两种高,表现为差异极显著(P<0.01)。三种试剂盒操作方法改进后提取的RNA经RT-PCR扩增猪-βactin基因,电泳结果显示,扩增条带清晰、特异,说明提取的RNA完全符合后续的分子生物学实验要求。Three commercial total RNA extraction kits. TRIzol, V-gene and H. Q. ＆. Q were applied to isolate high pure and integral RNA from adipose. It was found that it was hard to obtain high quality RNA when restrictively following their protocols. But with some modification, satisfied results were achieved. The purities of resulted RNA were not different among three modified methods. While the yield of modified TRIzol was higher when compared with other two. RT-PCR was used to detect the quality of RNA extracted by modified methods. Agarose gel electrophoresis showed clear, specific β-actin gene bands. It was proved that with these modifications, the pure and integral RNA could be obtained, which was suitable for common molecular biological researches.

关 键 词：脂肪组织 RNA提取 TRIzol方法 V-gene方法 H.Q.&.Q方法 RNA提取方法 脂肪组织 总RNA提取 操作方法 操作步骤 试剂盒 PCR扩增 分子生物学 提取过程 

分 类 号：Q522[生物学—生物化学]