Journal: Journal of Virology
Article Title: Natural 2?,5?-Phosphodiester Bonds Found at the Ligation Sites of Peach Latent Mosaic Viroid
Figure Lengend Snippet: Autoradiogram of 5% PAGE analysis of self-cleavage experiments using either 2′,5′- or 3′,5′-C-PLMVd. Radioactive transcripts (C-PLMVd) were added to the ground leaf powder, and the RNA was extracted using the RNeasy plant minikit and incubated under self-cleavage conditions with or without prior heat denaturation and snap-cooling treatments. Lanes 1 and 6, untreated L-PLMVd (control); lanes 2 to 5 and 7 to 10, 3′,5′- and 2′,5′-C-PLMVd, respectively; lanes 2 and 7, C-PLMVd extracted and incubated under self-cleavage conditions without heat denaturation and snap-cooling steps; lanes 3 and 8, like lanes 2 and 7, except that the extraction was performed in the presence of additional 5 mM EDTA in all buffers; lanes 4 and 9, like lanes 2 and 7, except that the samples were heat denatured and snap-cooled prior to incubation under self-cleavage conditions; lanes 5 and 10, like lanes 4 and 9, except that the extraction was performed in the presence of additional 5 mM EDTA. Adjacent to the gel, the positions of the C-PLMVd and L-PLMVd transcripts are used as size references.
Article Snippet: The first procedure used an RNeasy plant minikit (Qiagen) to prepare RNA from 50 to 100 mg of tissue as specified by the manufacturer.
Techniques: Polyacrylamide Gel Electrophoresis, Incubation