Enrichment of viroid small RNAs by hybridization selection using biotinylated RNA transcripts to analyze viroid induced RNA silencing
Enrichment of viroid small RNAs by hybridization selection using biotinylated RNA transcripts to analyze viroid induced RNA silencing
复制标题
DOI：
10.1007/s10327-008-0084-6
复制
发表时间：
2008-06-01
影响因子：
1.2
通讯作者：
Sano, Teruo
中科院分区：
文献类型：
--
作者：
Machida, Satoru;Shibuya, Makoto;Sano, Teruo
我们成功地应用了生物素化类病毒转录本和链霉亲和素偶联磁珠的杂交选择方法，然后再扩增、限制性内切酶酶切和连接，克隆了类病毒小RNA。用PSTVd特异的小RNA克隆效率比随机克隆未选择的小RNA高100多倍。该方法可用于分析小RNA的生物发生，特别是在各种宿主-病原体相互作用中寻找热点。
We have successfully applied a hybridization selection method using biotinylated-viroid transcripts and streptavidin-coupled magnetic beads, followed by re-amplification, restriction enzyme digestion and concatemerization, to clone viroid small RNAs. The cloning efficiency with the PSTVd-specific small RNAs increased more than 100-fold over that with random cloning of unselected small RNAs. The method can be used to analyze small RNA biogenesis, especially to find hotspots, in various host-pathogen interactions.