本研究利用CTAB法提取感染枣疯病的枣树叶片总DNA,再通过氯化铯-双苯酰亚胺密度梯度离心从总DNA中富集纯化枣疯病植原体DNA。利用枣树26S r DNA基因和枣疯病植原体16S r DNA基因的特异引物,分别对富集纯化前后感病枣树叶片总DNA进行PCR...本研究利用CTAB法提取感染枣疯病的枣树叶片总DNA,再通过氯化铯-双苯酰亚胺密度梯度离心从总DNA中富集纯化枣疯病植原体DNA。利用枣树26S r DNA基因和枣疯病植原体16S r DNA基因的特异引物,分别对富集纯化前后感病枣树叶片总DNA进行PCR扩增。结果表明,模板浓度为1 ng/μL时,富集后的总DNA模板扩增枣树26S r DNA的电泳条带亮度低于富集前,扩增枣疯病植原体16S r DNA电泳条带亮度明显高于富集前。Real-time PCR检测结果表明,模板浓度为1 ng/μL时,纯化后的枣疯病植原体DNA中26S r DNA相对纯化前的量为0.0638,而JWB相对纯化前的量为0.5035,这说明采用氯化铯-双苯酰亚胺密度梯度离心法可以有效地从感染枣疯病枣树总DNA中富集纯化枣疯病植原体DNA。展开更多
We present a systematic study of the effects of surfactants in the separation of single-walled carbon nanotubes (SWNTs) by density gradient ultracentrifugation (DGU). Through analysis of the buoyant densities, lay...We present a systematic study of the effects of surfactants in the separation of single-walled carbon nanotubes (SWNTs) by density gradient ultracentrifugation (DGU). Through analysis of the buoyant densities, layer positions, and optical absorbance spectra of SWNT separations using the bile salt sodium deoxycholate (DOC) and the anionic salt sodium dodecyl sulfate (SDS), we clarify the roles and interactions of these two surfactants in yielding different DGU outcomes. The separation mechanism described here can also help in designing new DGU experiments by qualitatively predicting outcomes of different starting recipes, improving the efficacy of DGU and simplifying post-DGU fractionation.展开更多
文摘本研究利用CTAB法提取感染枣疯病的枣树叶片总DNA,再通过氯化铯-双苯酰亚胺密度梯度离心从总DNA中富集纯化枣疯病植原体DNA。利用枣树26S r DNA基因和枣疯病植原体16S r DNA基因的特异引物,分别对富集纯化前后感病枣树叶片总DNA进行PCR扩增。结果表明,模板浓度为1 ng/μL时,富集后的总DNA模板扩增枣树26S r DNA的电泳条带亮度低于富集前,扩增枣疯病植原体16S r DNA电泳条带亮度明显高于富集前。Real-time PCR检测结果表明,模板浓度为1 ng/μL时,纯化后的枣疯病植原体DNA中26S r DNA相对纯化前的量为0.0638,而JWB相对纯化前的量为0.5035,这说明采用氯化铯-双苯酰亚胺密度梯度离心法可以有效地从感染枣疯病枣树总DNA中富集纯化枣疯病植原体DNA。
基金Acknowledgements Part of this work was financially supported by Grant- in-Aid for Scientific Research (No. 22226006 and 19054003), "Development of Nanoelectronic Device Technology" of New Energy and Industrial Technology Development Organization (NEDO), and the Global Centers of Excellence (COE) Program "Global Center for Excellence for Mechanical Systems Innovation". P. Z. acknowledges a scholarship granted by the China Scholarship Council and G. L. acknowledges support from the NanoJapan program funded by the National Science Foundation.
文摘We present a systematic study of the effects of surfactants in the separation of single-walled carbon nanotubes (SWNTs) by density gradient ultracentrifugation (DGU). Through analysis of the buoyant densities, layer positions, and optical absorbance spectra of SWNT separations using the bile salt sodium deoxycholate (DOC) and the anionic salt sodium dodecyl sulfate (SDS), we clarify the roles and interactions of these two surfactants in yielding different DGU outcomes. The separation mechanism described here can also help in designing new DGU experiments by qualitatively predicting outcomes of different starting recipes, improving the efficacy of DGU and simplifying post-DGU fractionation.