Abstract
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.
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Acknowledgments
This work was supported in part by Grants-in-Aid for Scientific Research C15580030 and B18380028 from the Ministry of Education, Science, Sports, and Culture of Japan, and a Grand-in-Aid for Scientific Research of Priority Area, Hirosaki University 2003–2006.
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Machida, S., Shibuya, M. & Sano, T. Enrichment of viroid small RNAs by hybridization selection using biotinylated RNA transcripts to analyze viroid induced RNA silencing. J Gen Plant Pathol 74, 203–207 (2008). https://doi.org/10.1007/s10327-008-0084-6
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DOI: https://doi.org/10.1007/s10327-008-0084-6