Establishment of the total RNA extraction system for lily bulbs with abundant polysaccharides
Establishment of the total RNA extraction system for lily bulbs with abundant polysaccharides
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富含多糖的百合球茎总RNA提取体系的建立
DOI：
10.5897/ajb10.2523
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发表时间：
2011-12-07
影响因子：
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通讯作者：
Li, Tianlai
中科院分区：
文献类型：
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作者：
Li, Xueyan;Wang, Chunxia;Li, Tianlai
百合鳞茎中含有丰富的多糖，因此很难从其鳞茎中提取高质量的RNA。为了解决这一问题，通过对十六烷基三甲基溴化铵(CTAB)、十二烷基硫酸钠(十二烷基硫酸钠)和Trizol法三种常见的RNA提取方法的改进，建立了一种高效、稳定的从百合鳞茎不同部位提取RNA的方法。为了去除鳞片中的多糖，提高了CTAB和十二烷基硫酸钠的氯化钠浓度。琼脂糖凝胶电泳和紫外光谱分析表明，所得RNA质量较高。此外，中间尺度的RNA产量最高，外部尺度次之，内部尺度次之。然而，Trizol法也进行了改进，以去除多糖，但失败了。逆转录-聚合酶链式反应(RT-PCR)结果表明，只有改良的CTAB法才能适用于后续的逆转录-聚合酶链式反应和文库构建。
Isolating high quality RNA from lily bulbs is made difficult by the presence of plentiful polysaccharides. To solve this problem, an efficient and steady RNA extraction technique from different parts of scales in three hybrids lily bulbs has been developed through modifications of three common RNA extraction methods, cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS) and Trizol method. NaCl concentration of both CTAB and SDS protocols was increased in order to get rid of the polysaccharides in scales. The resulting RNA is of high-quality, as judged by agarose gel electrophoresis and UV-spectroscopic analysis. Furthermore, the middle scales obtain the highest RNA yield followed by the exterior and the interior scales. However, Trizol method was also improved to remove polysaccharides but failed. The results of reverse-transcription-polymerase chain reaction (RTPCR) indicated that only the improved CTAB method is suitable for subsequent purposes, such as reverse transcription PCR and cDNA library construction.