| 罗辉,朱立成.紫藤种子总RNA提取方法研究[J].井冈山大学自然版,2017,(6):35-37 |
| 紫藤种子总RNA提取方法研究 |
| THE EXTRACTION METHOD OF TOTAL RNA FROM THE SEEDS OF WISTERIA SINENSIS |
| 投稿时间：2017-09-16 修订日期：2017-10-20 |
| DOI：10.3969/j.issn.1674-8085.2017.06.008 |
| 中文关键词: 紫藤种子 RNA提取 防御素基因 |
| 英文关键词: seeds of Wisteria sinensis RNA extraction defensin gene |
| 基金项目:国家自然科学基金项目（31360211，31460488）；江西省自然科学基金项目（20151BAB204020） |
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| 中文摘要: |
| 分离提取高质量的RNA是基因表达、调控和基因工程等研究的基础。植物组织中含有大量RNA酶、多糖和酚类等杂质，影响了高质量总RNA的获得。应用改良的CTAB法提取紫藤种子中的总RNA，添加了PVP、β-巯基乙醇和水饱和酚氯仿后能有效地除去大部分杂质，获得高质量的紫藤种子总RNA。3'RACE实验结果表明，以此总RNA反转录获得的cDNA为模板，可成功扩增紫藤种子中的防御素基因。 |
| 英文摘要: |
| Extracting high-quality total RNA from biological samples is the basis of investigation for the research of gene expression, regulation and genetic engineering. High levels of RNase, polysaccharides and polyphenol compounds make isolation of high quality total RNA difficult. In this study, a modified CTAB method was employed to extract the total RNA from seeds of Wisteria sinensis. After PVP, β-mercaptoethanol and phenol/chloroform were added, most of the impurities were removed effectively, and high-quality total RNA was obtained. The results of 3' RACE indicated that the gene of defensin in the seeds of W. sinensis was amplified successfully using the cDNA as a template which was got by RT-PCR from extracted total RNA. |
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