摘要
目的建立一种提取脂肪组织总RNA并能够有效地进行反转录-聚合酶链反应(RT-PCR)的方法。方法利用Trizol试剂提取人皮下脂肪组织总RNA,用紫外分光光度法鉴定其产率及纯度,用琼脂糖凝胶电泳和RT-PCR鉴定所提取的总RNA的完整性。结果采用选择性mRNART-PCR试剂盒特异性地扩增出脂联素(adiponectin)和β肌动蛋白(β-actin)的mRNA。结论在脂肪组织总RNA抽提中容易混入基因组DNA,而采用选择性mRNART-PCR只针对mRNA进行特异的扩增。
Objective In order to establish a method to extract total RNA from human adipocyte and effectively perform reverse transcriptase polymerase chain reaction(RT-PCR).Methods Using Trizol reagent to extract total RNA from human subcutaneous fat tissue.Ultraviolet spectrophotometry was used to determine extraction rate and purity,agarose gel electrophoresis and RT-PCR to identify the integrity of extracted total RNA.Result Using the RT-PCR kit containing dNTP analog can specifcally amplify the mRNA of adiponectin and β-actin.Conclusion The extraction of total RNA from fat tissue is often contaminated by genome DNA,but this defect can be readily overcome by the use of RT-PCR kit containing dNTP analog.
出处
《中国药物与临床》
CAS
2005年第4期265-268,共4页
Chinese Remedies & Clinics
