XUAN Jiping,JIA Zhanhui,QIAN Meihua,et al.Comparative of Methods for RNA Extraction From Plum (Prunus salicina Lindl.) Fruit Flesh[J].Northern Horticulture,2015,39(22):106-110.[doi:10.11937/bfyy.201522028]
李果实果肉组织RNA提取方法的比较分析
- Title:
- Comparative of Methods for RNA Extraction From Plum (Prunus salicina Lindl.) Fruit Flesh
- Keywords:
- plum (Prunus salicina Lindl.); fruit flesh; comparative; RNA; modified CTAB
- 文献标志码:
- A
- 摘要:
- 以中国李品种‘槜李’的果肉组织为试材，比较了Trizol〖KG-*3〗法、RNA提取试剂盒和改良的CTAB法提取RNA的效果，筛选出适合李果实果肉组织总RNA的提取方法。结果表明：Trizol法和RNA 提取试剂盒均不能得到完整的RNA；改良的CTAB法能有效去除多糖，28S条带的荧光亮度是18S条带的1.5~2.0倍，D260/D280值都在1.8~2.1，样品的平均得率为770.96~1 029.04 ng/μL，RT-PCR结果扩增出了特异性条带，说明改良CTAB法从李果实果肉组织中提取的RNA质量高、完整性好、产率高，完全可以用于后续的分子生物学试验。
- Abstract:
- To select the suitable method for total RNA extraction from oriental plum(Prunus salicina Lindl.) fruit，the performances of three extraction methods，trizol，plant total RNA extraction kit and modified CTAB were compared.Trizol and RNA plant extraction kit were determined to hardly extract integrate RNA.Modified CTAB was capable of efficiently removing polysaccharides.The lightness of 28S rRNA of the RNA isolated by modified CTAB was 1.5-2.0 times as much as that of 18S rRNA，with the OD260/OD280 value ranging from 1.8 to 2.1.The RNA productivities from 5 samples of fruit tissue were 770.96-1 029.04 ng/μL，respectively.The specific RNA bands obtained by RT-PCR indicate that the RNA extracted by modified CTAB was of high-quality and good-integrity at a higher productivity，thus completely suitable for further molecular biological research.
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备注/Memo
第一作者简介：宣继萍（1976-），女，浙江东阳人，博士，副研究员，现主要从事李果实发育等研究工作。E-mail：13915954315@163.com.责任作者：郭忠仁（1960-），男，江苏南京人，硕士，研究员，现主要从事果树育种和观赏园林植物资源等研究工作。E-mail：zhongrenguo@cnbg.net.基金项目：公益性行业（农业）科研专项经费资助项目（201003058）；江苏省农业三新工程资助项目（SXGC[2013]348）。