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当前位置: 首页 > 文章 > 青天葵叶片总RNA提取方法的比较研究 北方园艺 2013 (2) 99-102
Position: Home > Articles > Comparison of Total RNA Extraction Methods from Nervilia fordii(Hance) Schltr. Leaves Northern Horticulture 2013 (2) 99-102

青天葵叶片总RNA提取方法的比较研究

作  者：
梁凌玲;邓宇星;黄琼林;何瑞;詹若挺
单  位：
广州中医药大学中药资源科学与工程研究中心;广州中医药大学中药学院
关键词：
青天葵;总RNA提取;RT-PCR
摘  要：
采用RNeasy Plant Mini Kit法、RNAiso Plus法、RNAiso for Polysaccharide-rich PlantTissue法、CTAB法和SDS法,并结合4种抽提试剂组合,分别提取青天葵叶片总RNA。通过琼脂糖凝胶电泳、紫外检测及RT-PCR反应对提取效果进行了比较分析。结果表明:除RNAiso Plus法外,其它方法提取的总RNA均有一定程度的DNA污染。RNAiso Plus法组③(1/5体积KAC+4/5体积PCI)完整性最好,SDS法组④(1/5体积NaCl+4/5体积PCI)浓度最高,为372.4ng/μL。5种方法共20个处理组获得的RNA反转录后均能扩增出18SrRNA基因片段。该研究建立起了适用于青天葵叶片的总RNA提取方法,为青天葵的基因克隆、表达及转录组分析奠定了技术基础。
译  名：
Comparison of Total RNA Extraction Methods from Nervilia fordii(Hance) Schltr. Leaves
作  者：
LIANG Ling-ling1,DENG Yu-xing2,HUANG Qiong-lin1,HE Rui1,ZHAN Ruo-ting1(1.Key Laboratory of Chinese Medicinal Resources from Lingnan,Ministry of Education,Research Center of Chinese Medicinal Resource Science and Engineering,Guangzhou University of Traditional Chinese Medicine,Guangzhou,Guangdong 510006;2.School of Chinese Materia Medica,Guangzhou University of Traditional Chinese Medicine,Guangzhou,Guangdong 510006)
关键词：
Nervilia fordii(Hance) Schltr.;total RNA extraction;RT-PCR
摘  要：
Five different extraction methods(RNeasy Blant Miri kit,RNAiso Plus,RNAiso for Polysaccharide-rich Plant Tissue,CTAB and SDS),combined with four different extraction solutions were used to extract total RNA from Nervilia fordii(Hance) Schltr.leaves.The effect and efficiency of extraction was evaluated by agarose gel electrophoresis,ultraviolet spectrophotometry and RT-PCR.The results showed that apart from RNAiso Plus method,total RNA resulted from the other methods all showed genomic DNA pollution.Among these methods,the total RNA resulted from RNAiso Plus method followed with extraction solution ③(1/5KAC+4/5PCI) was the most integrated,and the total RNA resulted from SDS method followed with extraction solution ④(1/5NaCl+4/5PCI) had the highest concentration(372.4 ng/μL).Total RNA obtained from twenty treatment groups have been used to successfully amplify 18s rRNA gene segment by RT-PCR.This study established a proper total RNA extraction method for Nervilia fordii(Hance) Schltr.leaves,and it provided technique basis for gene cloning,expression,and transcriptomic analysis of the plant.

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