| 张锐,周开兵.荔枝果皮总RNA提取方法的筛选[J].中国南方果树,2013,42(5): |
| 荔枝果皮总RNA提取方法的筛选 |
| Study on the screening of the methods for the total RNA extraction from Litchi pericarps |
| 投稿时间：2013-08-03 修订日期：2013-09-16 |
| DOI： |
| 中文关键词: 荔枝 果皮 总RNA 提取方法 |
| 英文关键词:Litchi pericarps total RNA extraction method |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 中文摘要: |
| 以三月红荔枝果皮为试验材料，通过采用改良CTAB法、SDS法、改良SDS法、改良Bugos法和RNAplant plus Reagent试剂盒（TIANGEN公司）提取总RNA，采用浓度检测、纯度检测、RT-PCR等方法进行提取结果检测，进而筛选出最佳的提取方法。结果表明，这五种方法都能从三月红荔枝果皮中提取出总RNA，但总RNA浓度和纯度有明显的差异。其中采用改良Bugos法、改良SDS法和RNAplant plus Reagen试剂盒提取RNA的浓度较高；采用改良CTAB法和RNAplant plus Reagen试剂盒提取的总RNA纯度较高，完整性较好。采用SDS法、改良SDS法和改良Bugos法所提取的荔枝果皮总RNA纯度低，降解严重。经RT-PCR验证，采用改良CTAB法和RNAplant plus Reagen试剂盒均能扩增得到200bp的18sRNA编码基因片段。综合以上各项检测的结果，改良CTAB法为提取荔枝果皮总RNA的最佳方法，总RNA质量和得率满足后续分子生物学实验的需要。 |
| 英文摘要: |
| It is conducted with Sanyuehong litchi pericarps in Improved CTAB method, SDS method, Improved SDS method, Improved Bugos method and TIANGEN company’s RNAplant plus Reagent kit to research the screening of these methods through comparing the effects with each other by detecting concentrations and purities and RT-PCR. The total RNA from Sanyuehong litchi pericarps can be extracted in all these 5 methods, and the concentrations and purities of the total RNA are obviously different from each other among these methods. The concentrations of the total RNA extracted in SDS method and Improved SDS method and Improved Bugos method are higher. The purities and integrities of the total RNA extracted in Improved CTAB method and TIANGEN company’s RNAplant plus Reagent kit are higher and better, and those of the total RNA extracted in SDS method and Improved SDS method and Improved Bugos method are lower and worse. The both of the templates RNA abstracted in Improved CTAB method and RNAplant plus Reagent kit can be used for amplifying the fragment of 18s gene in RT-PCR. The quality of the total RNA extracted in the method of Improved CTAB in this paper is the best, and the total RNA is is fit for the subsequent operations such as reverse transcription and PCR and the construction of cDNA library. |
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