Analysis of small interfering RNA by capillary electrophoresis in hydroxyethylcellulose solutions
Analysis of small interfering RNA by capillary electrophoresis in hydroxyethylcellulose solutions
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DOI：
10.1002/elps.201500018
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发表时间：
2015-07
期刊：
影响因子：
2.9
通讯作者：
Chenchen Liu;Y. Yamaguchi;Xifang Zhu;Zhenqing Li;Yi Ni;X. Dou
中科院分区：
文献类型：
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作者：
Chenchen Liu;Y. Yamaguchi;Xifang Zhu;Zhenqing Li;Yi Ni;X. Dou
小干扰RNA （siRNA）的分析对基因功能研究和药物开发具有重要意义。采用CE法研究了siRNA阶梯标记在不同浓度和分子量（Mws）的羟乙基纤维素（HEC）聚合物溶液中的分离。siRNA阶梯标记是10个长度在20 ~ 1000 bp之间的双链RNA片段。通过迁移率和分辨长度（RL）图研究了dsRNA在CE过程中的迁移机制。我们发现RL不仅与HEC的浓度有关，还与HEC的分子量有关。例如，小dsRNA片段的RL受高Mw HEC浓度的影响大于大dsRNA片段，而大dsRNA片段的RL受低Mw HEC浓度的影响大于小dsRNA片段。此外，我们发现电泳证据表明，dsRNA的结构比相同长度的dsDNA更紧凑。在实践中，我们在4分钟内成功地从siRNA阶梯标记混合物中分离出了甘油醛3 -磷酸脱氢酶siRNA。
The analysis of small interfering RNA (siRNA) is important for gene function studies and drug developments. We employed CE to study the separation of siRNA ladder marker, which were ten double‐stranded RNA (dsRNA) fragments ranged from 20 to 1000 bp, in solutions of hydroxyethylcellulose (HEC) polymer with different concentrations and molecular weights (Mws). Migration mechanism of dsRNA during CE was studied by the mobility and resolution length (RL) plots. We found that the RL depended on not only the concentration of HEC, but also the Mw of HEC. For instance, RL of small dsRNA fragment was more influenced by concentration of high Mw HEC than large dsRNA fragment and RL of large dsRNA fragment was more influenced by concentration of low Mw HEC than small dsRNA fragment. In addition, we found electrophoretic evidence that the structure of dsRNA was more compact than dsDNA with the same length. In practice, we succeeded to separate the glyceraldehyde 3‐phosphate dehydrogenase siRNA in the mixture of the siRNA ladder marker within 4 min.