Enrichment of viroid small RNAs by hybridization selection using biotinylated RNA transcripts to analyze viroid induced RNA silencing

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 sm...

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Bibliographic Details
Published inJournal of general plant pathology : JGPP Vol. 74; no. 3; pp. 203 - 207
Main Authors Machida, S.(Hirosaki Univ., Aomori (Japan). Faculty of Agriculture and Life Science), Shibuya, M, Sano, T
Format Journal Article
LanguageEnglish
Published Japan Springer Japan 01.06.2008
Springer Nature B.V
Subjects
Agriculture
ARN
BIOCHEMICAL ENGINEERING
Biomedical and Life Sciences
Botany
Cloning
ENZIMA DE RESTRICCION
ENZYME DE RESTRICTION
GENIE BIOCHIMIQUE
HIBRIDACION
HOST PARASITE RELATIONS
HYBRIDATION
HYBRIDIZATION
INGENIERIA BIOQUIMICA
Life Sciences
Magnetism
Microbiology
Plant Pathology
RELACIONES HUESPED PARASITO
RELATION HOTE PARASITE
RESTRICTION ENZYMES
RNA
Technique
VIROIDE
VIROIDES
VIROIDS
RNA silencing
Hybridization selection
Small RNA
Viroid
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Summary: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.
Bibliography:2008006841
H20
ISSN:1345-2630
1610-739X
DOI:10.1007/s10327-008-0084-6